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Elements of applied microscop

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BOUGHT WITH THE INCOME OF THE SAGE ENDOWMENT FUND GIVEN IN 1891 BY

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ELEMENTS
OF

APPLIED MICROSCOPY.
A TEXT-BOOK FOR BEGINNERS.

CHARLES-EDWARD AMORY "^INSLOW,

Instructor in Industrial Microscopy

and Sanitary

Biology in the

Massachusetts Institute of Technology

FIRST EDITION.
FIRST THOUSAND.

NEW YORK:
London:

;:;

JOHN WILEY & SONS. CHAPMAN & HALL, Limited.'"^ <

1905.

I/t:

Copyright, 1905,

BY

CHARLES-EDWARD AMORY WINSLOW.

ROBERT DRUMMOND, PRINTER, NEW YORK,

DEDIgXTfiD

BY THE'!iUTHOR Zo Ibts /JBotber.

INTRODUCTION.

This

little

book

is

intended for the teacher, and the

It

beginner with the microscope, not for the specialist.

contains very few original data
ject with completeness.

and

treats

no

single sub-

Almost

all

the branches of technical microscopy have

been already
the present

made

the basis for

monographs with which

volume can in no way compete.

On

the

other hand, there are

many

entertaining popular bocks

it

on the microscope which

treat

as a

mere adjunct

to the

study of natural history.

Neither type of work was

suited for the use of a class in Industrial

Microscopy
Biologists

which
at

is

offered to second-year Chemists

and

the

Massachusetts
is

Institute

of
^first,

Technology.

The

object of this course

twofold

to give facility in

the manipulation of the microscope;

and second,
its

to fur-

nish an acquaintance with

.application.

the

scope of

practical

As a

text-book there was needed a brief

and elementary

treatise

which should take up the fundaitself

mentals of the science and art of microscopy

and

make a rapid but wide survey

which the

of the principal fields in

microscope has been

applied

to

practical

VI

INTRODUCTION.

affairs.
.

No

such elementary but comprehensive work

exists in

English

among

the

numerous able

treatises

upon
is

special branches of the subject.

The

present volume

therefore the outgrowth of a pedagogic need.

is

The book
but
if
it

necessarily incomplete

from the standpoint of the

it

expert in any of the branches which

treats,

conveys to the student's mind such ah idea of the possible

applications of the

microscope in varied

fields

as shall
of

stimulate

him

eventually to the further exploration

it

some one

of them,

will

have served
this

its

purpose.
authorities

In the preparation of

volume,

the

quoted in connection with the various chapters have been

freely

drawn upon.

The author

further desires to ex-

press his grateful obhgation, to Dr. P. G. Stiles for the

preparation of original drawings;

and

for advice

and

assistance in regard to various portions of the manuscript

to Professor

Professor
fessor

W. T. Sedgwick, Professor Mary A. Willcox, H. M. Goodwin, Professor F. J. Moore, ProWarren, Dr. C. C. Simmons, Mr. A. E.
G. Woodman, Dr. E. L. Walker, and
A.

C. H.

Leach, Mr.

Miss A. F. Rogers.

Acknowledgments
either

are"

due to the authors and pubfor figures

lishers of the following

books

which have

been copied directly or redrawn

Deschanel, a.

p., and Everett, J. D. Elementary Treatise on Natural Philosophy. New York, Appleton & Co., 1894. Hager, H., and Mez, C. Das Mikroskop und seine Anwendung.

Berlin,

J.

Springer, 1899.

Carpenter, W.

B., and Dallinger, W. H. The Microscope and its Revelations. London, J. & A. Churchill, 1891. Gage, S. H. The Microscope. Ithaca, Comstock Publishing

Co., 1904.

INTRODUCTION.

vii

Bausch, E. Use and Care of the Microscope. Rochester, Bausch & Lomb Optical Co., 1902. SCHIMPER, A. F. W. Anleitung zur mikrosl<opischen Untersuchung der vegetabihschen Nahrungs- und Genusmittel.
Jena, G. Fischer, 1900.

Hassack,

C.

Wodurch unterscheiden
Klepzig, 1900.

sich die Textilfasern

.'

Leipzig-Gohlis, a.

Herzberg, W. Papierpriifung. Ultzman, R., und HOFMANN, K.

Berlin,
B.

J.

Springer, 1902.

Atlas der physiologischen

undpathologischenHarnsedimente. Wien, Braumiiller, 1872. An Atlas of Bacteriology. Slater, C, and Spitta, E. J. London and Philadelphia, J. B. Lippincott Co., 1898. Wesbrook, F. F. Report of the Minnesota State Board of
Health,
1

899-1900,
C.

Whipple, G.

The Microscopy

of Drinking-water.

New

York, John Wiley

Galton,

F.

Sons, 1899. Finger-print Directories.

&

London, Macmillan

&

Co., 189s.

Howell, W. H.
Frazer,
1

Philadelphia and London,

p.
Bibliotics.

An American W. B.
A

Text-book of Physiology. Saunders & Co., 1901.

J.

Philadelphia,

B.

Lippincott Co.,

90 1.
O,

Lehmann,
1891.

Die Krystallanalyse.

Leipzig,

W. Engelmann,

Clark, C. H. Practical Methods in Microscopy. Boston, D. C. Heath & Co., 1894. LUQUER, L. M. Minerals in Rock Sections. New York, D. Van Nostrand Co., 1898. Sauveur, A. The Constitution of Steel considered as an Alloy of Iron and Carbon. Technology Quarterly, 1898, p, 78.

CONTENTS.

PAGE
i

I.

Function and Parts of the Microscope

II.

Manipulation of the Microscope

23

III.

The Mounting and Preparation of Objects for

THE Microscope
38
52 58

IV.

Micrometry, and the Camera Lucida

V.
VI.
VII.
VIII.

The Microscopy

of the

Common Starches

Foods and Drugs and their Adulterants

69

The Examination The Microscopy The Microscope

of Textile Fibres

80
95

of Paper
in Medicine

IX.

AND Sanitation

104
"126
141 153
ix

X.
XI.
XII.

Forensic Microscopy

Microchemistry

Petrography and Metallography

LIST OF ILLUSTRATIONS.

FIG.
1.

PAGE
Section through the

Human Eye

2.

Illustration of Refraction

4
5

3. 4.
5.

Refraction by Prisms
Refractions in Prisms of Different Angles

Plano-convex Lens and

to the Principal

its

Principal Focus
to the Relation of

6
a Luminous Point 7

6.

Course of Rays according

Focus

7.

8.
9.

Formation of Image by Object outside Principal Focus " " " " " inside " "

8 9
11

Leeuwenhoek's Microscope
Hooke's Compound Microscope
Course of Rays in Compound Microscope

10.
1 1.

12
13

12.
13.

Chromatic Aberration
Achromatic Objective
Effect of

14
15

14. 15. 16.

Homogeneous Immersion The Microscope

Relation of Distance from Principal Focus to Size and Position of

16
18

Image

21

17. 18.
19.

Proper Position for Observer

20.
21.

Method of Inserting Objectives Abbe Condenser Refraction by Air-bubbles and Oil-drops

Disturbing Effect of the Cover-glass

24 26 28
30
32 33
35

22.

Micrometer Calipers

23. Curvature of the Field 24. Turntable

42
'

25.
26.
27.

Thoma Microtome
Camera Lucida Course of Rays
in the

48
55 56
si

Camera Lucida

xii

LIST

OF ILLUSTRATIONS.
5
'

28.

The Common

Starches

29. Microscopic Structure of Coffee 30. Microscopic Structure of 31.

Mustard

75

"
Flax-fibre

"

"Pepper

32. Cotton-fibre
33.

77 ^3 ^S

34. Hemp-fibre
35. Jute-fibre

86
^7

36.

Ramie

88
9i

37. Wool-fibre 38. Silk

39. Tracheid of a Conifer

40.

93 loo
101

"

"Birch

41.

CellsofStraw 42. Urinary Sediment in Catarrh of the Bladder " " " Acute Bright' s Disease 43.
44. Blood-cells

102
108

no
113 115 118 122

45. Malarial Parasite 46. Diphtheria Bacilli

47. Trichina 48. Micro-organisms of Drinking-water 49. Microspectroscope
50.
."

124
131

Spectra of Oxyhsemoglobin and Reduced Haemoglobin

51. Finger-print Patterns

134 136
139

52.

Minute Characteristics of Handwriting Lines

53. Crystals of 54.

Hydrous and Anhydrous Ferrous Chloride Crystals of Cesium Alum

Crystals of Calcium Sulphate

144
145
145

55

Zone between Chlorides of Silver and lodin 57. Diagram of a Nicol Prism
56. Contact
58. Interference

148
156
161

Figures

59.

Curve of

Solidification for Alloy of Silver

and Copper

164
167

60. Microscopic

Appearance of Steel

ELEMENTS OF APPLIED
MICROSCOPY.

CHAPTER

I.

FUNCTION AND PARTS OF THE MICROSCOPE.

I.

Normal Vision.

^In

the ordinary act of vision at least

two

distinct processes

may

be distinguished, the formation

upon the
seen,
their

retina (K, Fig. i) of a minute

image of the object

optic nerves

and the transformation, by the

end organs, of the energy
which we

and

of the light-waves thus

thrown upon the retinal surface into that form of nervous

activity
call sensation.

The

first

process
is

fol-

lows the simple laws of Optics, while the second

trolled

con-

by the

far

more complex chemical and physical

conditions which furnish the subject-matter for the sci-

ence of Physiology.
rectly related to the
is,

The

sensation experienced
retina;

is

di-

image lying upon the

that
corre-

to

an area of special illumination whose parts

spond to those of some object outside, from which the

illumination
is

derived.

The mind has

learned,

by

experience, to interpret a certain image upon the retina

'

ELEMENTS OF APPLIED MICROSCOPY.

perceived by the as corresponding to a definite object but in other senses. The image is in all cases inverted,

found that images upside down correspond Experience to objects which are really right side up.
infancy
it

is

Fig.

I.

Section through the

Human

Eye.

(After Everett-Deschanel.)

A. Cornea. B. Aqueous humor.

C. Pupil.

K. Retina.

M.
N.

D.

Iris.

E. Lens. F. Suspensory ligament. G. Accommodation muscle.

L. Vitreous humor. Optic nerve. Inferior rectus muscle. O. Superior P. Levator palpcbrae muscle,
'
"

H.
/.

Sclerotic coat. Choroid coat.

^'

Meibomian glands.

teaches, also, that

an image
a
given

of a certain size

produced
implies

by an
definite

object

at

distance

always

magnitude; and we are therefore able to judge

size of

something about the actual

size of its

an object from the

its

image and our knowledge of

remoteness.

FUNCTION AND PARTS OF THE MICROSCOPE.

2.

Function of the Microscope.

In order that a visual

the'

image
retina

may be

produced, a certain definite area of

must be stimulated, and objects whose images are

smaller than this

minimum

will ordinarily

be

invisible.

Normal

vision

is

therefore limited to fairly large objects

If,

near at hand.

however, the rays proceeding from

more remote, or
lens

smaller, objects can be collected

to

by a

and bent so as
seen.

produce a larger image, the objects

is

may be
of large

This end

attained by the telescope

and

the microscope,

the former producing enlarged images

objects, the latter

and remote

producing enlarged

images of near and minute objects.

3.

Laws

of

Refraction.

^The
is

formation

of

such

images by the microscope

dependent upon the fact

that rays of light in passing

of
different

from any medium

a

to

one

density

experience

change in direction,

unless
contact.

they impinge at right angles to the surface of

The

deflection thus

produced has been compared to

the alteration in the course of a

column

of troops

on passfield.

ing from a smooth parade-ground into a ploughed

If,

in Fig.

2)

abaj)^ represents a

body

of troops
it is

march-

ing in the direction indicated by the arrow,

that

apparent

the

men on
field

the right of the line will reach the

first

ploughed
tarded.
the

biajf^a,

and

will

be somewhat

re-

The

left of

the line will gain

upon

the right

and

column as a whole

will execute a partial right face.

field

In passing out of the

on the other

side,
first

the right

of the line will reach the

smooth ground

and gain

as

much

as

it

lost

before, the final result being that the

ELEMENTS OF APPLIED MICROSCOPY.

in

column proceeds

a direction parallel to that which

it

Originally pursued.

be noted that in passing into the region offering a direcgreater resistance the marching column takes up
It will

tion

more nearly

at right angles to the

it is

boundary of that
line

region; in other words,

deflected toward a

drawn
So

normally, or at right angles, to that boundary

{11^.

Fig. 2.

Illustration of Refraction.

(After

Hager-Mez.)

when a ray of dense medium

it is

light passes
it is

from a

less

dense to a more

always refracted toward the normal,

while in passing from a more dense to a less dense

medium

bent away from the normal.

Refraction in the

4.

Convex Lens.

ray of light

passing through a piece of glass with parallel sides will

simply be shifted

laterally, as in

our illustration of the

marching column.
ever,

In passing through a prism, how3.

a resultant bending ensues as shown in Fig.

FUNCTION AND PARTS OF THE MICROSCOPE.

Ray AB, bent toward

and, bent

the normal at B, turns to the right the

away from
still

normal
If

to

the surface, at C,

turns to the right

further.

two such prisms are

Fig.

3.

Refraction by Prisms.

placed base to base as in the figure, rays striking them, as

do the

lines

AB

and AB',
meet

will

be bent in opposite direc-

tions so that they will

at the point at the

D.

The amount

of

bending varies with the angle

apex of che prism.

As shown,

for example, in Fig. 4, the ray a'Vc'd' passing

Fio. 4.

Refraction in Prisms of Different Angles.

through a prism of wide angle will suffer greater deflection than the ray abed
is

which meets surfaces

to

which

it

more nearly normal.

ELEMENTS OF yiPPLIED MICROSCOPY.

made biconvex lens works roughly, as if it were about a up of an infinite number of prisms arranged axis of such Rays parallel to the principal central axis.

curvature of its a lens (the Hne joining the centers of directions toward two surfaces, will be bent from all

and the rays nearest the outer edge or periphery will be most bent, so that all will meet at one point, known as the Principal Focus or burning-point of
that
axis,

the lens

(o,

Fig. 5).

The

greater the curvature of the

>

Fig.

s-

Plano-convex Lens and

its

its Principal Focus. (After Hager-Mez.)

lens the nearer to

will
5.
lie.

optical center this principal focus

Formation of Images by the Convex Lens.

is

As

the principal focus of a biconvex lens

defined as the

point at which rays parallel to the principal axis meet,

it

is

obvious that rays proceeding from a radiant point

placed at the principal focus will be sent off on the other

side of the lens as parallel rays,

and and

will

meet
Fig.

to

form an
Rays
i

image only

at

an

infinite

distance
lens

{B,
its

6).

from a point between the

being
still

principal focus,

more

divergent, will not even be

made

parallelj

but will

still

continue to diverge after passing through


FUNCTION AND PARTS OF THE MICROSCOPE.
the lens, though of course less so than before {A
,

Fig. 6)

No

real

image can therefore be formed by an object

lying inside the principal focus.

side the principal focus

Rays from a point

initial

out-

have an

divergence so small

as to be entirely overcome

by the

lens.

Such rays (C,

Fig.

6.

Course of Rays according to the Relation of a LuMmous Point to the Principal Focus.

Fig. 6) will actually converge after passage through the lens

and

will

meet

at a

definite

point to form a real

image.
6.

Construction

of

Real and Virtual Images.

^The
very

construction of the image formed by any object

simple,
lens
if

is

only

its

relation to the principal focus of the

be determined.

The

course of two rays from any

point i^

known with

certainty.

The

ray parallel to the

principal axis will be so bent as to pass through the principal focus

on the opposite

side of the lens.

The

ray

passing through the optical center of the lens will not be

ELEMENTS OF APPLIED MICROSCOPY.

all,

deflected at

because

it

will cut surfaces

which are

paraUel to each other.

Where
lens

these

two rays meet the

image
is

of the point

must be formed.

Thus

in Fig. 7

shown a biconvex

the surfaces of

which have
lie

an equal curvature;
equal distances from

its its

principal foci, therefore,

center.

at

side the principal focus F.

The The image

object ah

lies outis

of the point a

Fig.

7.

^Formation of Image by Object outside the Principal Focus. (After Hager-Mez.)

line aa^
is

determined by the straight

flcfflj.

and the broken

similarly fixed,

line

The image
will

of the point h

and
a'h'.

between the two

This

will

be formed the enlarged image

be a real image

one, that

is,

which could be

caught upon a screen held in the right plane,

and

it

will

be inverted.

These are the

characteristics of all images

focus.

formed by objects lying outside the principal

Fig. 8 illustrates the other case, in
inside the principal focus.

which the object hes

it

Here

is

evident that the

rays from the point a will not meet at

side
of

all

on the opposite image can be

the

lens

and

therefore

no

real


FUNCTION AND PARTS OF THE MICROSCOPE.
formed.
point
9

They

will

appear to the eye to come from a

their

more remote than

from

true

position;

and the
This

exact location of this point

the lines

may be found by

prolonging-

and

F backward

until they meet.

Fig.

8.

Formation of Image by Object inside the Principal

Focus.
(After Hager-Mez.)

image

will not

be inverted, but erect

it is

and as

it

cannot be

caught upon a screen

7.

called a virtual image.

Development of the Simple Microscope.

The

effect

of globes of crystal in concentrating the sun's rays

single point

on a

was known

in very early times.

Aristophanes,

Pliny the Elder, and other Greek and

Roman

authors
Sen-

mention the use of such primitive burning-glasses.

eca states that "letters though small and indistinct are

seen enlarged and more distinct through a globe of glass
filled

with water."

None

of the ancients appear,

howthis

ever, to

have thought of any practical appUcation of

as an aid to vision;

phenomenon
up

and medical

writers

to the thirteenth century of the Christian era

speak of

short-sightedness as an incurable infirmity.

In microscopy, as in so

many

other branches of knowl-

lo

ELEMENTS OF APPLIED MICROSCOPY.

first

edge, the

distinct

advances were

made

in Arabia.

Alhazen, the great Arabian physician

of the eleventh

century, distinctly describes the use of lenses for pro-

ducing enlarged images, in a pubHcation supposed tc

date

from

about

1052.

Two

centuries

after,

Roger

Bacon, the Franciscan monk, and noted alchemist, of Oxford, noted the same effect. A little later, near the

end

of the thirteenth century, lenses

were

first

applied

to the mitigation of defects of vision;

and

to Salvino
of

d'Armato degli Armati, a Florentine, the invention

spectacles
is

ascribed.

The

use of lenses as microscopes

for the examination of objects too

minute to be studied

with the unaided eye became general about the end of

the sixteenth century. vations were numerous,

From
and

the year 1600 such obser-

in 1637 the first

diagram

of

a microscope

now

extant was published by Descartes.

About 1665 small

glass globules

began

to

be used instead

of convex lenses for the simple microscope.

set in

They were

metal plates, on the side of which opposite to the

observer the object to be examined was mounted on some

sort

of

movable arm.

With

these instruments a high

magnification was

obtainable;

and

it

was with such and Leeuwen-

simple microscopes (Fig. 9) that the pioneer microscopists

of the seventeenth century, Kircher in Italy

hoek
8.

in Holland,

founded the science of Micro-Biology.

Microscope.

The Compound

Meanwhile

step

had been taken, which, though not particularly fruitful at the time, was to become later of great significance.
This was the invention of the compound microscope

commonly

attributed

to

the

Dutch spectacle-makers,

FUNCTION AND PARTS OF THE MICROSCOPE.

n
from

Hans and Zacharias Janssen, and supposed

about 1590.
It is certain that

to date

compound microscope

was independently invented by Galileo in 16 10, and that

Cornelius

Drebbel in Holland was credited with the

introduction of the instrument about 1621.

These early

Fig. 9.

^LEEtrwENHOEK's

Microscope (Circa

1700).

(After Carpenter-Dallinger.)

instruments and their illuminating apparatus were cum-

brous and unwieldy in the extreme (see Fig. 10).

The compound microscope

that
it

is

characterized by the fact

contains two or more lenses or systems of lenses,

one of which forms an image of the object, while the other

forms a second image of the

the rays in such

first
is

image.

The

course of

an instrument

shown

in Fig. 11.

The

object .4 jB

lies

outside the principal focus of the lens system

its
is

marked Objective, and

at

real inverted

image

is

formed

A^B^.

This image

produced

at a point inside the

principal focus of the Ocular or Eyepiece;

and the

eye-


12

ELEMENTS OF ylPPLIED MICROSCOPY.

an erect and
virtual image,

piece forms

A'^B^ of the

real

inverted image
9.

A 'J5*.
Compound Microscopes.The com-

Defects of the

FiG. 10.

Hooke's Compound Microscope

(After Carpenter-Dallinger.)

(1665).

pound microscope was

tance,

at first of little practical impor-

on account

of

the complications introduced by

spherical

and chromatic aberration.

to the fact that the rays

The former is due which pass through the outej;

FUNCTION AND PARTS OF THE MICROSCOPE.

13

Fig.

II. Course or Rays in the Compound Microscope

(After Gage.)

14

ELEMENTS OF

/IPPLIED MICROSCOPY.

part or periphery of a biconcave lens are brought to a focus somewhat closer to the lens than those which lie nearer
the optic axis.
of the

This deflection and the consequent blurring

image increase with every increase in the curva-

ture of the lens.

still

more

serious defect lies in the

fact that the rays of light of different colors are differ-

ently affected

by the ordinary

lens, those of shorter

wave-

length at the violet end of the spectrum, {v), coming to a

focus

first,

as indicated in Fig. 12.

The

result

is

that


FUNCTION AND PARTS OF THE MICROSCOPE.
(in virtue of

IS

which white Hght

is

spHt up into

its

compo-

nents) varied independently of each other.

Thus flint glass

glass
If a

has only

.i

to .2

more

refractive

power than crown

with more than twice

lens of

its

dispersive power.

biconvex

crown

glass, (C),

be combined with a plano-con,

cave lens of
as

flint glass,

(F) arranged in the opposite sense,

possible so to adjust their oppoflint glass

shown

in Fig. 13,

it is

site
all

curvatures that the

shall

compensate
glass

for

the dispersion caused

by the crown

and neu-

FiG. 13.

The Achromatic
its

Objective.

(After Hager-Mez.)

tralize

only half

refraction.

This

is

the principle of
the
efforts

the

achromatic objective;

and through

of

SelUgues and Chevalier in France, Fraunhofer in Ger-

many, Amici
England,
it

in Italy,

and Goring, Tulley, and

its

Lister in

gradually attained

practical development

between 1820 and 1830.

With this improvement the compound microscope acquired new importance; and
very shortly developed into one of the most important

it

instruments at the disposal of modern science.

II.

The Immersion
flint

Objective.

Lenses

made

of

com-

bined crown and

glass in such fashion

as to be

achromatic will also show a decreased spherical aberration,

and

this latter defect

may be

further reduced to a

minimum by

adjusting the radii of curvature of the oppo-


i6

ELEMENTS Oh APPLIED MICROSCOPY.

surfaces of the lenses according to certain

site

known
in-

relations.

Such lenses are designated as aplanatic.

still

There

remained one serious limitation to the

crease of the

power
In

of magnification of the

compound
is

microscope.
of light,
ture,

all its ^^arious

refractions there

a loss

and with the small

objective lenses of great curvait is

necessary for high magnification,

difficult to get

a sufficient illumination for clear vision.

it

Furthermore,

has been shown by

Abbe and

others that the rays

which extend from a point toward the periphery of a

lens

are
finer

of

prime

importance
of
objects.

in
If

the

detection

of

the

structure
lost,

the

outer zones of
details

rays are

no image
this
is

of

very minute

can

be formed; and

just

what occurs

in
air

the ordi-

nary compound microscope

the objective

when
to

there

is

between
is

and the specimen

be examined, as
If,

shown

in the right half of Fig.

14.

on the other

Fig. 14.

Effect of Homogeneous Immersion.

(After Hager-Mez.)

hand, some substance

like cedar-oil,

which has the same

refractive index as glass,

be placed between the lens and

will

the cover-shp

which covers the specimen, the rays

FUNCTION /IND PARTS OF THE MICROSCOPE.

i?

take the direction indicated on the left-hand side, more

peripheral rays will enter the lens, and more detailed images
will

be produced.

Such an immersion objective was

first

suggested by Amici in 1850, and next to the achromatic

objective this
in

may be

considered the greatest single step

the improvement

of the

compound microscope.

The
of light

degree to which a lens admits the peripheral rays

is

designated by the term angular aperture, which

signifies the angle

contained between the most divergent

rays passing through the- objective from the axial part

of

an object

(a point situated

on the principal

axis of

the lens).

Obviously the angle will increase with the

its

convexity of the lens and

tance
;

consequent short focal

dis-

with the same lens, the angle will be greater

is

when
to

some homogeneous immersion substance

this

used.

Taking

factor into account, the

power

of

an objective

collect
ical

and

utilize divergent light-rays is called its

Numerby

Aperture;

this quantity is

equal to the index of re-

fraction of the

medium

in front of the lens multiplied

the sine of half the angle of aperture.

12.

The Mechanical Parts

its

of

the Microscope.

^The
and

microscope consists in
lenses of

essentials of the

two systems of

which we have spoken.

For steadiness these

rigid stand;
light

optical parts

must be riiounted upon a

is

in addition apparatus

needed for throwing

upon

the object to be

examined and

for focusing, or so ad-

justing the
that a clear

relation

between the object and the lenses

image may be produced.

illus-

microscope of the ordinary American pattern,

15,

trated in Fig.

has a heavy horseshoe- shaped

base

ELEMENTS OF APPLIED MICROSCOPY.

Fig. 15.

^The Microscope.
j^'
r

A. Base. B. Pillar.
C.

Coarse adjustment.

Arm.

L. Fine adjustment.

D. Tube.
E. Collar. F. Objectives. G. Ocular.

M.

Stage.
clips.

N. Spring
O. Mirror.

P. Mirror bar.
Q. Substage. S, Diaphragm.

H. Draw-tube scale. Z, Draw-tube.

FUNCTION AND PARTS OF THE MICROSCOPE.

and a
vertical pillar rising

19

from

it,

which may or may

the stage,

not be jointed.
perforated plate

Attached to the

pillar is

upon which the object may be placed;

Hghting mechanism, and above
is

below

this is the

the

arm

carrying the optical parts.

lighting apparatus includes
first

The

a mirror, usually

having one plane and one concave surface and so mounted

on a jointed bar as
it

to set at

any angle.
light

For high powers from the mirror

lens

is

customary to concentrate the

of

by means
below the

an Abbe condenser, a large

placed

stage.
itself

The

stage

in the simpler microscopes

in the center,

is

merely a

fiat plate with

an opening

and with clamps

is

for holding the, slide

upon which
is

the object

mounted.

For

delicate

work

it

convenient to use a microscope

equipped with a mechanical stage which can be moved

forward and backward and from side to side by a micrometer

screw.

Such an arrangement makes

it

easier

to

explore the whole of the specimen to be examined, and

enables one to find any particular portion of

ure.

it

at pleas-

Under
for

the stage, whether

it

be of the simple or
sort of dialight

the mechanical type, there should be some

phragm
will

regulating the

direction

from which

reach the object, and

cutting off the peripheral rays

itself

coming from outside the object

obscure the image.

which tend

to

The diaphragm opening must

there-

fore vary with the size of the object

under examination

and with the power

of the microscope, a smaller opening

being used with a higher power.

This adjustment

is

accomphshed by a revolving

disc with openings of dif-

ELEMENTS OF APPLlEb MICkOSCOPY.

and by an
Iris

ferent size in the cruder microscopes,

diaphragm

in
is

more

elaborate instruments.
pillar

The arm
moves
adjustment,

borne by the upper part of the

which

in the lower part,

is

and

this

motion, called the fine

regulated by a milled head at the top.

the distance between the lenses

By
sive

this

means

and the

object

may

be varied with

changes in the position

More extengreat delicacy. of the lenses are made possiwhich they are fixed sHdes
it

ble

by the

fact that the tube in

up and down
the arm.

in the

clamp which holds

at the

end

of

This movement, called the coarse adjustment,

may be regulated by hand or by a rack-and-pinion. The tube itself is divided into two portions, the
proper and an inner cylinder, the draw-tube, which

tube

may
Into

be pulled out telescope fashion from

its

upper end.

the lower end of the tube proper the various objectives

may be
is is

screwed, while the eyepieces or oculars

slip into

the upper end of the draw-tube.

When

the draw-tube

pulled out the space between the two systems of lenses

increased,

and

in order that the real

visible

image produced
it

by the objective may be

through the eyepiece,

must be formed farther up than would ordinarily be the

case.

A reference

to Fig. i6 will

show

that,

according to

the principles previously deduced, this can be accom-

plished

of the objective,

by bringing the object nearer the principal focus and that the image of the object so

brought nearer the principal focus will be correspondingly

enlarged.

Thus
sets

pulling

out

the

draw-tube

in-

creases the magnification.

At

least

two

of lenses

accompany a compound

FUNCTION AND PARTS OF THE MICROSCOPE.

microscope,

21

commonly two

eyepieces

and two or more

objectives of different magnifying powers.

The

lenses of

some makers are designated by

rule,

arbitrary symbols;

as a

however, they are marked with numerals which

indicate

what

is

known

as their equivalent focus, or the

focal length of a

simple converging lens which would

Fig. i6.- -Relation of

Distance erom Principai Focus to Size AND Position or Image.

size as that

produce an image of the same

the lens in question.
will
less

formed by
the lower

be the power.
than a

The larger the number Thus a ij-in. eyepiece

and a

magnifies

i-in. eyepiece,

J-in. objective less

than

a ^-in. objective.
tive is the tV>

The commonest high-power

is

objec-

and the ^^

the highest objective which

can be practically constructed.

violet rays

By making

use of ultra-

with a wave-length only half that of ordinary

light the Zeiss Optical

to construct

Company has recently been able The instruments of much higher power.
and the image
recorded on the
invisible,
is

lenses in this case are of fused quartz,

produced, being of course

photographic plate.

22

ELEMENTS OF APPLIED MICROSCOPY.

In some microscopes the two lenses of the eyepiece

work together

to

produce the

effect

shown

in Fig.

ii.
is

More commonly, however,

formed between these two
one forms the
virtual

the

real

inverted image

lenses,

and only the upper

an eyepiece,
ocular, the lower

image.

In such

known

as a negative or

Huyghenian

or field lens really forms part of the objective system.

REFERENCES.
Bausch, E. Manipulation of the Microscope. Rochester, 1901. Carpenter, W. B., and Dallestger, W. H. The Microscope and its Revelations. London, 1901. Clark, C. H. Practical Methods in Microscopy. Boston, 1894. Gage, S. H. The Microscope. Ithaca, N. Y., 1904. Hager, H., and Mez, C. Das Mikroskop und seine Anwendung.
Berlin, 1899.

These

references,

and those appended

to other chapters, include

only those few books which have been found most indispensable in
the work of the course in Industrial Microscopy as given at the Institute

of Technology.
will

very

full

bibliography of works on

microscopy

be found in Professor Gage's book cited above.

CHAPTER

II.

THE MANIPULATION OF THE MICROSCOPE.

I.

Setting

Up

the

Microscope.The microscope

is

an instrument
impaired by
it

of precision

whose dehcacy may be

easily

carelessness or neglect.

When
it

not in use
in
its

should be protected from dust by placing

case
settle

or

under a beU-glass, and any particles which

it

upon

should be removed with a camel's-hair brush

If

and chamois-skin.

necessary,

the mechanical parts

may be

first

cleaned with chamois-leather moistened in a

solution of equal parts of benzine

and

ohve-oil

and then
the base,

wiped with dry chamois.

The microscope should be handled always by

and not by
straining the fine adjustment.

the upper part of the piUar, in order to avoid

AVhen in use

it

should be

placed on the work-table rather near the edge, with the

pillar side nearest the observer,

who

should

sit

close to

the table in a chair of such height that his eyes will

little

be a

above the level of the upper end of the draw-tube

Fig.
17).
sit

(see

Those unfamiliar with the microscope

For general work
23

are apt to

too far off or too high above the instrument

it.

and bend painfully toward

best not to
tilt

it

is

the tube of the

microscope even when

the


24
pillar is

ELEMENTS OF APPLIED MICROSCOPY.

provided with a joint for that purpose;
is

and

if

the instrument

kept upright, the spring clips

may

be
the

dispensed with, the object being

stage as desired.

moved about on

Fig. 17.

^Proper Position for Observer.

(After Bausch.)

The

lenses of the microscope should always

be examthey
re-

ined before they are inserted in order to see

quire cleaning.
soft

if

Since the glass

still,

is

easily scratched, only

is

clean cloth or, better

Japanese lens-paper

suitable for this purpose;

the fingers, which are always

MANIPULATION OF THE MICROSCOPE.

oily,

25

must never be allowed

to

touch the

lens.

Well-

defined spots seen on looking through the microscope at

a clear field are due to specks of dust

as

upon the

eyepiece,

may be proved by

rotating that system of lenses.

Not
sur-

infrequently such specks are

formed on the upper

face of the lower or field lens of the ocular,

which must

be unscrewed in order to remove them.

ness
ive;
is

Diffuse cloudi-

generally caused
this
it

by

dirt or

moisture on the objectbreathing on the glass

and

may be removed by
with lens-paper.

and wiping
smears of

Sometimes for stubborn

alcohol or xylol

dirt or grease,

95%

may be
may
not

used, but in the

latter case the solvent

must be sparingly
it

appKed and promptly removed

affect the setting of the lenses.
2.

in order that

Lighting.

^After the objective

has been screwed in

18,

at the

bottom of the tube, as shown in Fig.

and the

eyepiece has been inserted at the upper end of the drawtube, the mirror should be so adjusted as to give the best

available illumination.

Direct sunlight must of course

fur-

be avoided, but a clear*area of sky or a white cloud

nishes,

an

ideal sourcfe of light;

and
For

sunlight reflected

from a white wall or transmitted

tain

directly through a curartificial

may be used

to advantage.

illumina-

tion electric bulbs with ground-glass shades are suitable,

or the narrow edge of the flame from a flat-wicked

oil-

lamp may be

substituted.

gas-flame,

unless

some
is

incandescent mantle be attached to the burner,

satisfactory

not

on account of

its

unsteadiness.

The

flat

surface of the mirror yields sufiicient light

under ordinary circumstances, but the concave surface

26

ELEMENTS OF /tPPLIED MICROSCOPY.

may be used when the strongest illumination is desirable. The observer should so adjust its angle in relation to the
window
light

or

lamp

that a clear

and well-defined

circle of

may

be seen.
if

If the

hght be uneven, or

an image of the window-bars

so changed as to re-

or the flame be visible, either the angle of the mirror or

its

distance from the stage

may be

o o

Fig. i8.

^Method or Inserting Objectives.

If the circle
is

(After Bausch.)

move

the difficulty.

too bright so that

its

glare hurts the eyes, the angle of the mirror

must be

changed

to

moderate

it

and the diaphragm opening

desirable that the light should

should be decreased.

For most purposes

pass directly

it is

upward through
This end

the axis of the microscope-

axial illumination.

may be
and

attained

by

focus-

ing just below an air-bubble,

so arranging the mirror

that the bright point inside shall be exactly at the center of the bubbleIn examining diatoms and some other

MANIPULATION OF THE MICROSCOPE.

27

objects, oblique illumination brings out certain structures

best.

Light so oblique that

it

cannot enter the objective

at all is

sometimes obtained by placing under the stage

off the

a stop cutting

central cone of light.

In such a

preparation the object will be

made

visible

by the rays
will

which

it

reflects

or refracts upward,

and

appear

self-luminous
illumination.

on

black

background

dark-ground
concave

With

objectives higher than \ in. .even the

.for

mirror will not give sufficient light

scopic work,

successful micro-

and the Abbe condenser must be used, with

the plane mirror.

The

condenser, as has been stated in

of lenses placed just
a'

Chapter

I,

is

an objective system

under the stage in such a position as to concentrate

considerable

amount

of light

upon the

object to be ex-

amined

(Fig. 19).

With low powers the condenser should

its

be swung out from

TV-in-

place to one side;

results

while with. the

immersion objective the best

oil

may be obtained

by placing a drop of
bottom
of the slide,

between the condenser and the

entire system optically

making the
one
is

homogeneous.
magnifications

When
it

working with these high

the condenser
that
is,

is

also important that

should be accurately centered and focussed;

it

must be

at

such a distance below the stage that the great-

est possible

amount

of light

the object.

This position

may be concentrated upon may be determined by focus|-in. objective

ing

upon some specimen with a

thfe

and then

so adjusting

condenser that the image of a windowitself.

sash or of a flame coincides with the object

It

has been pointed out that the function of the dia-


28

ELEMENTS OF APPLIED MICROSCOPY.

is

phragm
finally

the rays to cut off adventitious light so that all

reaching the

microscope

shall

come from

the

immediate

vicinity of the object.

A little

experimentation

will show that reducing the diaphragm opening often

gives a sharper picture as well as one

to the eyes; in general

much

less trying

front lens of

an opening about the size the objective will yield good results.

of the

Fig. 19.

Abbe Condenser.

(After Hager-Mez.)

Certain opaque objects are best examined by reflected

light,

and

in

many
no

cases this

method may well be used

to

supplement the ordinary one, the mirror being turned

aside so that
light passes

through the stage and the

object being illuminated either with the light which naturally falls
by.

on

it

or with rays concentrated from above

a lens or mirror.
Focusing.
is

3.

After
on the

the field

is

well lighted, the next

step

to focus

object, or to so adjust the relation

of the lens systems that a clear

image

may

be formed.

M/INIPUL/1TION OF THE MICROSCOPE.

29

The low-power
for preliminary

objectives

(those of one-half inch

and

one-third inch equivalent focus) should always be used

exploration before a

new specimen

is

examined with the higher powers, since

of

their large field

clear.

view makes the general relations of the object

is a,lso

Focusing

much

easier with the lower powers, since

with them a small object

their
tive

may

easily

be found, while

working distance, or the space between the objec-

and the object when

in focus,

is

great

enough

to

allow free play.

After placing the object over the center

of the opening of the stage, the tube should

be run down
is

by means

of the coarse

adjustment until the front lens

within an eighth of an inch of the object.

Then, looking

through the microscope, the tube

is

slowly raised

by the
less

coarse adjustment until the object comes more or

clearly into view.
It is often helpful to

move

the slide

about with the

left

hand

at the
is

same

time, as the

shadowy

outUne of a moving object

that of one at rest.

more

readily recognized than

As soon

as the object

is

dimly seen

it

clearly into focus

by the use
this

of the fine adjustment.

may be brought The

is

proper manipulation of
portance, since
if

mechanism

of great im-

the attempt be

made

to study objects

which are not well

in focus, the eyes will

be strained and

structures incorrectly seen.

Furthermore, at any given

position of the fine adjustment only a certain plane of the

object examined will be in focus, while

its

whole figure

is

needed to make up a correct picture.

Therefore, one
fine

hand should be kept continually on the

while the microscope
is

adjustment
needs

in use, varying

it

slightly as

ELEMENTS OF /tPPLIED MICROSCOPY.

It

demand.
fine

should be noted that the excursion of the

is

adjustment
it

necessarily small;
its

if

care
its

is

not taken
will

to

keep

near the middle of

it

range,

motion

be
its

stopped short at one end or

place at the other.

will

be unscrewed from

Focusing with the higher power

is

more
it

dif&cult.
is

On

account of the shprt working distance

necessary to

run the objective down as close to the object as possible,

watching
its

approach from the

side,

and then focusing

up

very carefully with the fine adjustment.

Useful practice in focusing study of

clove-oil

may

be obtained by the

air-bubbles

and

oil-drops.

few drops

of

should be mixed with mucilage and the mixture

beaten up on a slide with a knife-blade so as to insure

\J

MANIPUL/ITION OF THE MICROSCOPE.

the ring widening the
this

31

objective
is

passes

and the center becoming sharper as downward. With the oil-bubble

brightest

reversed, the

center
is

and the widest,

seen

ring being
Fig.
18.

above.

Why

this

so will be

from
less

The

oil-bubble,

a dense sphere in a

dense medium, acts hke an ordinary lens concentrating

light to

a bright focus above, the surrounding ring from

is

which the hght

will

diverted being dark.

This dark ring

widen upward.
less

The

air-bubble,

on the other hand,

being
light,

dense than the mucilage, diverges the rays of

4.

and its -dark ring is widest below. The Use of the Draw-tube. ^The

spherical

and

chromatic aberration of an objective vary with

tion to the object
fore

its rela-

examined and

to the eyepiece.

There-

when we

say that in an achromatic objective these

this is only true for

defects

have been corrected,

set of conditions.

a certain

standard

If

we change
certain

the position of

the draw-tube
objective

and

therefore the distance between the

and eyepiece systems, a

again introduced.

amount

of aberis

ration

is

For each objective there

certain proper tube-length, record of

which accompanies
objective

the microscope
itself.

and may be placed upon the

to

According

the

best

American practice the

of the tube

tube-length,

measured between the upper end

is

where the eyepiece

inserted
is

and

the lower
is

end

of the

tube where the objective

inserted,

either

160 or

216

mm.
cover-glass
is

A
and

commonly placed between

and

the object

and the objective

in order to hold the former in place

this introduces a pertur-

to protect the latter;

32

ELEMENTS OF APPLIED MICROSCOPY.

Reference to Fig.
cover-glass shifts
arise

bation which must be allowed for.

21 will

show

in a general

way how a

the rays coming

from a

point,

F, and makes them

apparently from F' and F".

In the making of achrotaken into

matic objectives this

account;

effect of the cover-glass is

but with each lens, standard results can only

be obtained with cover-glasses of a certain thickness.

variation of .05

mm.

in thickness

may

quite obliterate

Fig. 21.

^DiSTUEBiNG

Effect of the Cover-glass.

(After Gage.)

certain fine structures.

Adjustable objectives are so made

that

by turning a

ring or collar the distance between

their systems of lenses

may be
may
is

varied, being increased

for the thinner cover-glasses.

With an ordinary unadselect

justable objective one

cover-glasses

of the

thickness for which

easily

it

corrected, the dimension being

is

measured by some such apparatus, as

shown

in

Fig. 22.

Or

the lens system

may be
by

adjusted for cover-

glasses other than the standard

altering the position of

the

draw-tube,

changes in the

tube-length

producing

changes similar to those which are provided for in the adjustable objective.
for thin covers

The

tube-length should be increased

for those thicker than the

and decreased


M/INlPULy4T10N
standard.

OF THE MICROSCOPE.
in

33

Such changes

adjustment are necessary only

in the case of delicate work.

S.

Care of the Eyes.

Much
lenses,

of the difficulty some-

times experienced by beginners with the microscope

may

be avoided by attention to three points mentioned above

the cleanliness of the

illumination of the
the fine adjustment.

a clear but not excessive

of

field,

and a proper manipulation

In no case should the observer

strain his eyes in the attempt to study

what he cannot

Fig. 22.

^Micrometer Calipers.

see clearly.

If the object is not distinct, there

must be

something wrong which should be remedied.

The

distance of the eye above the eyepiece should

vary with the magnification.

lenses there
is

For each combination of

a certain eye-point at which a

is

maximum
and above

number
shadows

of rays

most

closely concentrated,

or below,' the size of the field will be reduced,

will appear.

and colored

The

higher the power the nearer

the eye-point approaches to the eyepiece.

It is best to

accustom oneself to the use of both eyes

to acquire the habit of keeping

alternately,

and

open the

eye which

is

not over the instrument.

At

first

the atten-

34
tion

ELEMENTS OF APPLIED MICROSCOPY.

may be

distracted

by external

objects, but this

diffi-

culty will pass with practice.

The beginner

may

also be

misled by seeing through the microscope certain cloudy

specks floating across the field of view.

These are

the

muscae volitantes, shreds of matter lying in the vitreous

humor

of the eye; after a time they are so discounted

their

by the Observer that he becomes unconscious of

presence.
6.

Qualifications of a

Good Microscope.

^The

princi-

pal parts of the

compound microscope have been

it

de-

scribed in Chapter I;
of the

remains only to point out some

most important

qualities

by which we may measure

Differences in

the value of any individual instrument.

the mechanical parts are largely a matter of personal

preference, although the stand should be as compact as
possible with a basa sufficiently heavy to give proper
steadiness.

Both the coarse and

fine

adjustments must

work
slip

easily without being so loose as to allow the tube to

down

of its

own

weight.

have an ample excursion.

The fine adjustment should The mirror-bar ought to

any position
in

move

freely,

and yet

retain

which

it

is

placed.

The

stage should be wide

enough

to

accommo-

date large objects

when

necessary.

The
of

optical parts are, of course, of prime importance;

they should be examined with respect to the four qualities

magnifying power, resolving power, penetration, and

illuminating power.
ratio

The

magnification of a lens, or the

size of the
its

between the

size of

an object and the

image

formed, depends simply on the curvature of

surfaces

and may be measured by the method described

in Chap-


MANIPULATION OF THE MICROSCOPE.
ter

35

IV.

The

resolving

power by which

fine structures are

made
as has
tested

visible varies directly

with the numerical aperture,

been already explained.

This power

is

ordinarily

by examining the wing-scales of certain Lepidopthe

shells-

tera or

of diatoms which
shell of

possess very fine

markings.

For example, the

Pleurosigma shows

three systems of striations

nification

when examined under a maglesser aper-

of

250

diameters with a numerical aperture

of over .80, while

ture.

two of them disappear with

Penetration, or the

power

to see clearly different

planes of the object at the

same time,

varies inversely

with the numerical aperture and directly with the square

of the equivalent focus of the lens.

Illuminating power

varies with

the square of

the numerical aperture,

and

with the square of the equivalent focus.

Two common

faults should

be looked for in a microcorrection of

scope, curvature of the field

and imperfect
If

chromatic and spherical aberration.

ter,

a stage microme-

marked with

lines at right angles,

a, Fig. 23.

be examined, the
it

image should resemble

If

has the appear-

a
Fio. 23.

b
(After Hager-Mez.)

Cttrvature of the Field.

ance of 6 or

the lenses are so ground that the magnifica-

tion is greater or less at the periphery than at the center.

36

ELEMENTS OF APPLIED MICROSCOPY.

Spherical aberration
of the test objects

may be

detected by examining one

If the lens

mentioned above.

be under-

corrected, so that peripheral rays

come

to

a focus nearer

the lens, the outer part of the object will be in focus in a

plane below the central portion.

Chromatic aberration

may

be detected in the examination of any object by the

it.

appearance of colored rings surrounding

7. Interpretation of Appearances.

The

greatest care
to

must be taken
structure

in

drawing conclusions as

the real

of bodies

from

their

appearance under the

microscope.

Right and

left

are reversed by the invert-

ing action of the objective lenses.

Appearances in any
air-

one plane

may be
its

very deceptive, as in the case of an

bubble at
object
as
is

upper edge.

The medium
III.

in

which an

mounted may completely

in

alter its appearance,

we shall see membered that

Chapter

It

must always be

re-

contrasts of density are

what give us mi-

croscopic pictures;

only

when

these coincide with salient

differences in structure will such pictures be representative.

The

presence of dust or other foreign materials

may

mislead the observer, and even air-bubbles have

If the preparation

sometimes caused confusion.

treated with fixing agents,

has been

dyes,

etc.,

as described in
to mistake

Chapter
artificial

III, great care

must be exercised not

conditions due to reagents for those normally

existing.

Motion
deceptive.

of bodies

under the microscope

is

is

particularly

Since the distance travelled

magnified, an

idea of rapid
translation

movement

is

conveyed when the actual

Diffusion currents

may

be really very slow.

MANIPUUTION OF THE MICROSCOPE.

are often set

37

up

in the

mounting medium which may be

misinterpreted.
in the

still

more

serious source of error lies

phenomenon known

as the

Brownian movement,
affects

or pedesis, a dancing, oscillating motion which

suspensions of finely divided soHd particles under certain not very clearly understood conditions.

Gamboge
Brownian

or carmine

suspended

in

water shows

the

movement
that such
teristic of

well,

and should be studied

carefully in order

motion

may

not be confused with that charac-

the bacteria or other living organisms.

REFERENCES.
Batjsch, E.

Manipulation of the Microscope.

Rochester, igoi.

Gage,

S.

H.

The

Microscope.

Ithaca,

N. Y., 1904.

CHAPTER

III.

MOUNTING AND PREPARATION OF OBJECTS FOR THE MICROSCOPE.

I.

The

Effect

of

Mounting Media.

^The
Under

clearness

with which

we

see

an object depends
its
its

in great part

upon

the contrast between

density or color

and the backthe mi-

ground furnished by
croscope
little

surroundings.

remains but difference in density to empha-

size outlines,

and the

distinctness of a given object will

thus vary widely according to the

lies.

medium

in

which

it

The

student

may

obtain an idea of the importance

it

of this factor

and

of the necessity for taking

into ac-

count in interpreting microscopic appearances, by the

examination of potato-starch grains in

cerin,

air,

water, gly-

and

clove-oil.
little

In

air the grains will

show heavy

black edges and

internal structure.

In water the

edges are less pronounced and the hilum and oystershell

markings appear.

In glycerin the same characin clove-oil the

teristics are still

more pronounced, and

edges are almost invisible, so that the grains have a spectral

appearance and the hila are very strongly marked.

it

Obviously,

is

necessary in making deductions as to

the real structure of an object to consider the influence

38

MOUNTING
of the

/IND PREP/IR/tTION

OF OBJECTS.
also desirable

39

mounting medium;

and

it

is

by

choosing a

medium
is

of proper density to

minimize as far

as possible the errors

due

to excessive refraction.

Thus,

when
tive

starch
is

mounted

in air, the difference in refrac-

Index

so great that the heavy black edges produced

is

are most deceptive, while in clove-oil the difference

slight for clear definition.
2.

too

Temporary Mounting Media.

it

When

an object

is

of such nature that

can be well examined in

air, it-Kiay

be placed upon an ordinary glass

I in.,

slide (usually 3 in.

by

best with ground-glass edges)

and examined

directly
if

under the microscope.

Even

in this case, however,

the high objectives are used, the specimen should be

covered with a cover-slip in order to protect the front lens.

Of

the cover-glasses

commonly

sold the square ones are

most convenient for ordinary handling, and the round

ones best suited for making permanent mounts, as will

be seen

later.

The

effect of the thickness of the cover-

shp upon microscopic vision has already been considered in Chapter II.

In

fine

work

it is

often desirable to
select

measure the thickness of the covers used in order to

those of a standard size or to adjust the objective or

draw-tube to those which deviate from

it.

This

may
upon

be effected

conveniently by means

of micrometer calipers

or with certain special forms of apparatus placed

the

market for

this

purpose (Fig. 22).

in in

Most objects are more clearly seen medium than air, and, when mounted
cover-slip
is

some denser
any
liquid,

always necessary even for low powers.

The

choice of a

medium must be determined by two

consider-

40
ations.

ELEMENTS OF APPLIED MICROSCOPY.

First,

as suggested above,

its

refractive

index

should be far enough, and yet not too

that of the object to be examined.
of such a nature that
it

far,

removed from
it

Second,

should be

will not set

up

destructive, os-

motic, chemical, or other changes in the substance of the

specimen.

For aquatic plants and animals and a

fulfils

great

many

other objects water

these requirements; and

for specimens of animal tissue the

normal

fluids of the

body furnish
jects,

ideal
it

mounting media.

With denser

ob-

where

is

necessary to minimize contrast

effects,

glycerin or

some

oil is preferable.

3. Principles

of

Permanent Mounting.

^In

making
necessary

permanent mounts of microscopic objects

to

it is

guard against changes due to

of the object,

(a) physical displace(6)

ment
ical

mounting medium, or cover;

chem-

changes in the substance or the mounting medium or

loss of the latter

by evaporation; and
its

(c)

bacterial decomFirst, then,

position of the specimen or

the cover-glass

substratum.

must be firmly attached

to the slide either

by an adhesive mounting medium or by the

of a special cement.

application
pro-

Second, the

medium must be

tected that
it

from evaporation and made of such a nature

will not

undergo chemical change.

Third, the

mounting, medium must be antiseptic.

These requirements are

ent ways:

fulfilled in three quite differ-

by mounting "dry" or

in air, in

which case

the absence of moisture serves as an antiseptic,

by mount-

ing in aqueous media mixed with glycerin, and by mounting in balsam, a resin which sets and forms a solid mass

impervious to

all

external agencies.

MOUNTING
4.

y4ND PREP/IRATION

OF OBJECTS.
of

41

Mounting Dry or in Air.^The method

is

dry

mounting

well suited to such objects as crystals, which

show well

in air,

and which may be dried without

It
is

suf-

fering alteration.

fairly

simple, although in this

it

respect, as well as in

permanency,

is

excelled

by the
cleaned

balsam mount.

For
slides

all

methods

of

mounting,
first

thoroughly
Slides

and covers form a

essential.

which

have been already used and are soiled with balsam or

other resinous substances must
or turpentine.
first

be treated with xylol

Otherwise

all

slides

and covers should

of water
this they

be put into a cleaning mixture made up by dissolving

20 grams of potassium bichromate in 100
cc.

and adding 100

cc.

of sulphuric acid.

From

may be

transferred to

50%

alcohol,

and

in that solution

kept until needed for use.

Both

slides

and covers should

oils of

always be handled by their edges, since the

skin will infallibly soil their surfaces.

the

Given the
structed

slide

and the

cover,

there

must be conshall separate

some

sort of wall or cell

which

the two

and prevent the specimen from being crushed.

vary in depth with the thickness of the

The

cell will

object, and,

sary to
directly

when a shallow cell will serve, it is only necesmake a ring of cement and place a round cover upon it. A neat mount may be made by using
is

a turntable such as

shown

in Fig. 24, the slide being

clamped on the movable portion and whirled about,

while a brush

dipped in shellac, marine glue.

is

Bell's

cement, or some similar material

the guide rings

held just over one of

tip lightly touch-

on the turntable with


42

ELEMENTS OF

yIP PLIED

MICROSCOPY.

ing the glass.

narrow, even ring

may

thus be pro-

duced, and, by a gradual process of upbuilding, one of

considerable depth.
to If

an object of greater thickness

is

be examined, a ring of brass or hard rubber

may be

attached to the slide by a ring of cement similarly applied.

When

the cell has

become so dry

that there

is

no dan-

ger of the cement running, the object to be mounted

may

be placed within

it

or attached to the under surface of a

Fig. 24.

^Turntable.

(After Gage.)

cover-slip.

The

cover

is

then pressed gently

is

down on
all

the ring of cement so that contact

complete

the

way round,

and, after placing the slide on the turntable

is

once more, a fresh ring of cement

the edge of the cover

apphed
slide.
is

partly on

and

partly

When

thus prepared the

on the dry mount

completed and
slide,
it

should be labelled on the left-hand end of the

the

nature of the specimen, the treatment to which

has

been subjected, the mounting medium, and the date being

indicated.
S.

Mounting in Glycerin Media.

Glycerin

mounts,
bal-

although not easy to

make and

less

permanent than

MOUNTING
sam mounts,
removing
all

/IND PREPAR/ITION
suitable
it

OF

OBJECTS.

43

are

for

microscopic plants and

other objects which

is

desirable to examine without

the water
is

which they contain.

Some
placed

pre-

hminary treatment
cells

necessary even in this case, since

containing a large

amount

of water,

if

di-

rectly in

a strong solution of glycerin, would be torn

and

distorted

by the

violent diffusion currents set up.

is

The
to im-

most convenient way to prepare such objects

merse them in

io%

glycerin

and

set

them by

for a

few

days under cover, as a protection from dust, so that the water

may

evaporate and produce a gradual concentra-

tion of the glycerin.

Specimens thus prepared

cell filled

may be mounted
it is

in a

deep

with pure glycerin, but

difficult

matter to

cover the cell so that air-bubbles shall not be included,

and a

still

more

difficult

matter to cement the cover

firmly to the wetted surface of the ring.

For most

pursince

poses glycerin jelly *

this

is

much more
cement

convenient,

substance becomes solid on cooling and obviates

to attach the cover-

the necessity for a cell or for

slip.

All that
it

is

necessary in using this

medium

is

to

melt

by gentle warmth, cover the object with a goodit,

sized drop of

put on the cover-slip, and

set aside in

cool place.
6.

Mounting

in

Balsam Media.

The most permanent

is

and

satisfactory of all

mounting media
is

Canada balsam,

* One part of gelatin by weight

7 parts of pure glycerin are

soaked in 6 parts of water

finally

added and

i%

of phenol.

The

mixture
till it is

is

warmed
and
is

for ten to fifteen minutes, with constant stirring,

clear

then

filtered.

44

ELEMENTS OF y^PPLIED MICROSCOPY.

and has a high index
distinct.

since this substance sets like a rock

of refraction,
It is

which makes opaque objects very

commonly used

dissolved in xylol, the natural fir

oil;

balsam being mixed with an equal volume of the

the solution after filtering
is

concentrated to a syrupy
It
is

consistency
jelly,

by evaporation.

used

like

glycerin
set aside,

being placed on the object, covered and

the
xylol

when

evaporates

and the balsam becomes

firmly set.

With an object
wing or

of firm texture, not containing water,

the preparation of a balsam

leg of

mount

is

very simple.

The
be

an

insect,

for example,

may

easily

mounted
those
to

in this way.

With

softer specimens, especially

it

made up

largely of water,

is

necessary to resort
shall

some preliminary treatment which

tissues.

harden and

dehydrate the
7.

Fixing and Dehydration.

The

first

step in the prep-

aration of a soft plant or animal tissue for balsam mount-

ing

is

to treat

it

with some agent which shall

fix

the

cell

structures in exactly the condition in

in
life,

which they occurred

preventing the disintegrating changes which norshall

mally follow the death of protoplasm, and which

at the

same time so

act chemically

upon the

cell constitu-

ents as to harden

and protect them from the action

later.

of

the

chemical agents to be applied

is

Flemming's
it

mixture
tains

one of the best of these fixing agents;

acid,

con-

.25% chromic

.1% osmic

acetic acid dissolved in water,

acid, .1% glacial and should be allowed to

act for half

an hour or more.
is

saturated solution of

corrosive

subUmate

often

useful.

Absolute alcohol

MOUNTING AND PREPARATION OF may be

applied to the fixation of

OBJECTS.

45

many

tissues, its action

being so rapid as to forestall the bad effect of the violent

dehydration which

it

sets

up.

At the other extreme,

dilute alcohol (i part

90%

alcohol in 2 of water) pro-

duces

moderate

fixation

without

harmful

osmotic

changes.
After fixing with

weak

alcohol, or- after

washing out

Flemming's
next step
is

fluid or corrosive sublimate

with water, the

to dehydrate,

which can best be done by

increasing
strength.

treatment

with

alcohol

of

After

the specimen has been thoroughly permeated with

alcohol,
to
is
it

30%

should be transferred to a

50%

solution, then

70%, 90%, and 95%, removed so gradually

successively.

Thus

the water

that the diffusion currents set

up

are not sufficiently violent to distort the tissue.

The

period of immersion in each grade of alcohol will vary

with the thickness of the

specimen.

For

sections or

minute objects three to

five

minutes will

suffice.
is

The
to treat

last step
it

before mounting an object in balsam

is,

with a clearing agent, that

with some liquid

of high refractive index

which
as

will penetrate its tissues

and make them

grains

clear, just

glycerin

makes

starchin
air.

more transparent than when mounted

is

Since the object

in

already in alcohol and

is

to

be mounted

balsam,

it is

obvious that the clearing agent should be

Cedar-oil, clove-oil,

miscible with both substances.

xylol are perhaps the

and

commonest

clearing agents.
it is

After

being treated with any one of them until

transparent, the object

thoroughly

may be

placed in balsam and

mounted.

46

ELEMENTS OF ylPPLIED MICROSCOPY.

The

clearing agents just described

depend

for their

action solely

upon the

fact that their high index of re-

fraction prevents loss of the light passing through the

object.
ical

In the preparation of certain very opaque botanare

specimens, clearing solutions

used which act

dissolve
cer-

chemically
tain

upon

the tissues

and actually

constituents which

interfere with transparency.

Of

these,

strong

aqueous solutions of potassium hydrate

or chloral hydrate are

commonly

used,

and they must

be allowed to -act upon the specimen for several hours or

even days.
8.

Section-cutting.
entire,

Since only very small objects can

necessary in

be examined

it is

many

cases to pre-

pare thin sections of objects for examination with the

microscope.

Even with

fibres

and

similar objects which

can be easily studied in one dimension, cross-sections are

often desirable in order to gain an idea of their whole
structure.

Sections of rigid objects

may be

cut directly with a

sufi&ciently

sharp razor;

but few specimens are

this

hard

to

be treated in

way.

In general, plant and animal

tissues are so soft that they

would give way even before

a sharp knife, and must be supported by imbedding

them
to

in

some material

of firmer texture.
thin,
it

If the sections

be cut are not very

is

only necessary to surlike pith;

round the specimen as a whole by a substance

if

more

delicate

work

is

to

be done, the tissue must be

like

permeated by some material

paraffin
cell

or celloidin

which
it

will

support each individual

wall and

make

rigid.

MOUNTING AND PREPARATION OF

OBJECTS.

4ll

Plant stems and leaves, of which sections a tenth of a

miUimeter or more in thickness will serve,

may be

easily

imbedded and cut

in

pith.
is

The

fresh tissue, without

slit

preliminary treatment,

placed in a

cut at

theend

of

a section of moistened elder pith.

the pith, a the pith
left
is

A thread is tied around

hold
all

little

below the end,

to

together.

Then

taken between the thumb and forefinger of the

hand, with the forefinger almost level with the end

and the thumb lower down.

the right hand,

The

knife or razor, held in

forefinger of the left
to cut off a thin

and steadied on the

hand,

is

drawn toward the body so as

its

sKce of the pith with

tion thus cut

imbedded specimen.

The

sec-

may

then be examined at once in water or

prepared in dilute glycerin for a glycerin mount, or

passed through the grades of alcohol for the balsam mount.

When

thin sections,

down

to thousandths of

milli-

meter in thickness, are desired, the object

must be im-

bedded in paraffin or
this
is

celloidin

and

cut with a microtome

tissues,

almost
softer

always

necessary with animal

tissues,

which are

than plant

and

at the

same time
the object

more opaque.
must
first

For imbedding

in

paraffin,

be

fixed,

dehydrated, and cleared, as described

above.

It is

then placed in paraffin, kept at a temperaits

ture just above

melting-point in a suitably regulated

to

bath,

and allowed

remain until thoroughly saturated.

to

The

liquid parafl&n
sort of

and the object are then transferred

cell

some

temporary
it

made

of paper, or of metal

blocks, in

which

can be cooled rapidly by a current of

crystals of paraf-

"running water.
fin fatal to

Slow cooling produces

clean sections.


48

ELEMENTS OF /IPPLIED MICROSCOPY.

Once
well imbedded, a cube of solid paraf&n containis

ing the object

as
is

cut out

and

fixed in

a microtome, such

illustrated in Fig. 25.

Several good instruments are

on the market,
Blake types

of

which the

Thoma and

the Minot-

may

be mentioned.

In any case the knife

and

object are firmly held in supports

moving

at right
is

angles to each other;

and one or other of the supports

Fig. 25.

Thoma Microtome.

(After Carpenter-Dallinger.)

SO arranged that
shifted

by some mechanical device

it

can be

by any desired amount between each

excursion,
If the

according to the thickness of the sections desired.

surrounding temperature be right, the successive sections

of paraffin will adhere

by

their edges,

forming a long

ribbon.

In such a ribbon each

serial section represents

one plane in the original specimen; and the whole object

may

thus be unrolled as

it

were upon the

slide,

each

structure being easily traced

from section

to section.

Minute sections obviously cannot be handled by themselves.

It

is

desirable, therefore,

to affix

them

to the


MOUNTING ANO PREP/IRATION OF
slide

OBJECTS.

49
this is
fixa-

upon which they are

to

be mounted;
of Mayer's

and

best

accompUshed by the use

albumin

tive.*

A minute

drop of

this solution is
till

placed on a clean
only the thinnest

sUde and rubbed with the finger

film remains.

The

section
till

is

then placed upon the slide

and gently heated

the paraffin just melts.

The

sec-

tion will adhere firmly to the

shde and the paraffin

may
unfor

be dissolved in xylol or any other clearing agent.

For objects over lo

suitable, since

mm.

in diameter paraffin
split

is

large blocks

under

the knife;

such
ding
ether,

specimens celloidin

may be
is

used as an imbed-

medium.
and

This

substance

used

dissolved

in

after the evaporation of the latter is

hardened
process
it

by treatment with alcohol or chloroform.

is less

The

simple than the paraffin procedure, and

is

not

possible to cut such thin sections of the objects

imbedded.

The

freezing microtome, in

which the water in the

Its

specimen acts at a low temperature, as

substance,
is

imbedding
Cell

rapid and yields very thin sections.

structures are, however,

ess,

somewhat

distorted in this proc-

and

its

use

is

confined mainly to the preparation of

pathological material.
9.

Staining.
for

One more

process in the preparation of

still

objects

the

microscope

demands reference
Since the elements
it is

the process of differential staining.

of a tissue or of a cell differ in

chemical composition,
shall

possible

to

apply

certain

dyes which

enter into

combination with some of them and not with others,

* 50
erin
cc.
I

of the

albumin of hen's eggs

of

is

mixed with 50

cc. of glyc-

and

gram

sodium

salicylate,

shaken well and

filtered.

50

ELEMENTS OF APPLIED MICROSCOPY.

their

and thus emphasize

ture

outhne and bring out the

stain
acts

struc-'

more

clearly.

Sometimes the
it

upon

certain tissue as a whole, picking

out from other tissues

surrounding

it.

More

generally, however, the substances'

cell,

used act on a certain part of the individual

the

generally

nucleus,

making

that

structure

stand out clearly

from the cytoplasm.

Staining processes are often complicated,

and
is

include,

not only direct methods in which the stain

act just long

allowed to

enough

to affect the desired elements

and
thc'

then washed out, but also indirect methods in which

tissue is overstained

and the dye then removed by alcohol 5r acid from the parts which give it up most readily.;
Small objects
treated on the

may be
slide.

stained

in

bulk and sections

Of
anilin

the

staining

solutions

used,

haematein

and

the

dyes

(fuchsin,
etc.)

methylene blue, eosin, safranin,

are the

Bismarck brown,

most generally
it is

useful.

By

a proper combination of two of these

possible to

stain the cytoplasm of the cell

one color and the nuclei

of

another.

This process of double staining depends

course on the greater avidity of the nuclear substance

for

most dyes.
is

good example for practice in double

staining
tion,

the haematein-eosin combination.

The

sec-

attached to the slide with albumin fixative and

is

cleared,

passed
is

down through

the grades of alcohol to

solu-

30%, and

then placed for one or two minutes in a

tion of Mayer's haemalum.*

The
in
cc.

section

is

then washed
alcohol with

Mix

gram haematein dissolved

50

cc. of

90%

50 grams alum dissolved in 1000

water.

Cool, settle,

and

filter

MOUNTING
in

/IND PREP /I RATION

is

OF OBJECTS.

5'

water

till

the stain

removed from the cytoplasm

and passed through 30%, 50%, and

this stage
it is

70%

alcohol.

At

immersed

for a

few seconds in a solution

of eosin in

70%

alcohol to stain the cytoplasm,

and

is

then passed rapidly

up through 90% and 95% alcohol

and

xylol to

be mounted in balsam.

The

nuclei should

be dark blue and the cytoplasm reddish yellow.

Another good stain for the beginner to use, though not

to

make

up,

is

the Wright modification of Leishman's

blood-stain,

which with a single solution yields a .most

beautiful differentiation of half a dozen cell elements.

Its use is described

more

fully in

Chapter IX.

REFERENCES.
Chamberlain, C. J. Methods Gage, S. H. The Microscope.
Lee, a. B.
in Plant Histology.

Chicago, 1961.

Ithaca,

N. Y., 1904.
Philadelphia, igoo.

The

Microtomists'

Vade Mecum.

ZiMMEEMANN, A.
trans.

Botanical Microtechnique.

HuMpheey,

J. E.,

New

York, 1893.

CHAPTER

IV.

MICROMETRY AND THE CAMERA LUCmA.

I.

The Stage Micrometer.

whose spread

Objects

may

be

directly

measured under the simple microscope by the use

dividers,
is

of

determined, under the

lens,

by means
meter.

of a steel scale divided into fifths of a millispecial standard is

lines

With higher powers, some

and
.i

required;
intervals of

glass slides bearing fine

.01

ruled at

and

mm.

are prepared for this pvurpose.

These are known as stage micrometers.

,

The
the

imit in
is

which the measurement of microscopic objects

com-

monly expressed

is

.001

mm., known as

microletter
/t.

miUimeter or micron, and denoted by the Greek

Obviously such a scale
for
is

the only absolute essential

micrometry or measurement with the microscope,

it

and

would be possible
it

to find the dimensions of

an

object by simply placing

ofif

on the ruled

slide

and reading
is,

the

number
open
to

of spaces covered.

This process
Accurately

how-

ever,

two

objections.

graduated

stage micrometers, which are

expensive, would be too

their

subject to breakage to

make

employment

in this
it is

manner economical.

Furthermore, on such a scale

58

MICROMETRY
difficult to

/IND

THE CAMERA LUCIDA.

since, as

S3

measure small objects,

lie

mounted on
and

the slide, they tend to

their edges

obliquely across the scale,

rarely

would only
therefore,

coincide with

its

lines.

As a
ment
2.

rule,
is

some

indirect

method
most

of measure-

adopted.

The Ocular Micrometer.

of

The

satisfactory

method

measurement with the microscope involves

the use of another scale, called the ocular micrometer.

This

is

a circular piece of glass cut to

fit

in the eyepiece

of the
at

microscope and bearing a

scale

made up

of lines

an equal, but not necessarily a known, distance apart.

is

The micrometer
resting

to

be placed inside the eyepiece,

inside
it,

on the

sliding

diaphragm
lie

which should
image

be so adjusted as to

in the plane of the real

formed by the objective.

the scale,

Thus

'the

image
it.

will lie

on

and may be measured by

The
of

value ob-

served

is

therefore the size of the virtual

image of an

object formed
scale.

by the objective in terms

an arbitrary

In order to obtain actual dimensions the ocular

micrometer must be standardized in order to determine

the value of
its

divisions as
size.

compared
this

to the
it is

image of an
simply nec-

object of
essary to

known

For

purpose

place a stage micrometer under the microscope

it

and focus upon

with the ocular micrometer in position.

The

ratio of ocular divisions to stage divisions (or rather

to the

images of stage divisions) can then be read

off

and, knowing the actual value of the stage divisions,

easy to calculate the

it is

number

of microns

on the stage

which correspond to one ocular division.

the ocular micrometer
is

When

once

standardized for a given system

54
of lenses

ELEMENTS OF APPLIED MICROSCOPY.

and length
of draw-tube, the stage micrometer

may be

discarded.
it

In using the ocular micrometer

have seen, that the diaphragm

is

necessary, as

we
be

in the eyepiece should

at a proper level or the scale will not

be

distinct.

Too
scale

strong a light also blurs the image of the micrometer

divisions.

The

object

to

be measured and the

should be arranged in relation to each other, by rotating

the eyepiece

and adjusting the

slide, so that

one edge of

the object shall coincide with the edge of one of the lines
of the scale.
ness, care

Since these lines have a considerable thickto refer the object to correits

must be taken

sponding edges of the lines at

extremities.

The

diffi-

culty of estimating the boundaries of objects with accu-

racy

is

always great

and the

limits of precision of meas-

urement
3.

in the best microscopic

work

is

about

.2

fi.

Measurement with

the
of

Camera

Lucida.

Any
will

method by which the image

an object may be com-

pared with the image of a scale of known dimensions

serve for

measurement with the microscope

is

and, although
satisfac-

the ocular micrometer

the simplest

and most

tory process, there are several others, of which one involv-

ing the use of the camera lucida

tioned.

may

be

briefly

menis

This apparatus, whose external appearance

in Fig. 26,
fits

shown

over the eyepiece of the microscope.

reflect the rays of

It contains
light

a system of mirrors which

coming from a paper

laid beside the microscope, so

that they enter the eye along with the rays

which pass
the

up through the microscope.

object

Thus

the

image of

upon the

stage

and

that of the paper are super-


MICROMETRY /tND THE C^MER/I LUCIDA.
posed one upon the other.
matically
Fig.

55

27 indicates diagram-

the

general arrangement of the mechanisni.

is

At P, just over the eyepiece,

a cube

made up

of

two triangular prisms of glass with a silvered surface

between them pierced by a central opening.

this

Through
from

opening the rays from the microscope pass, while

those

coming from the paper, AB, are

reflected first

the swinging mirror at the side

and then upward from

Fig. 26.

Camera Lucida.

(After Gage.)

the silvered surface of the prisms.

distortion
"angle
it

In order to avoid

is

necessary that the mirror should be at

surface
this

an

of 45" to the

on which the paper

1

rests.

In order to bring

about

it

is

necessary to sup-

port the paper oh an inclined surface

the microscope extends

when

the base of

outward so as

to interfere

with a

viewof the

table.

The adjustment

of light with the

difficulty.

camera lucida
the
light

is

matter of considerable

If

passing

through the microscope be too bright, only the object will


56

ELEMENTS OF APPLIED MICROSCOPY.

if,

be seen, and
the paper
invisible.
is

as

more commonly
the illumination

occurs, the light from

the strongest, the object becomes almost

is

When

properly balanced,

both object and paper should be clearly seen, and the

point of a pencil on the paper
outline of

may be made
accuracy.

to trace the
this

the

object

with

Obviously

Fig. 27.

Course of Rays in the Camera Lucida.

(After Gage.)

device, primarily intended as

an aid to the making of

drawings of microscopic objects,

etry.

may be

used in microm-

With a given arrangement

drawing on the paper
will

of apparatus, the size

size of

of the

depend upon the

the object.

By removing

the latter
will

and

substituting a

stage micrometer

whose image

be superposed upon

that of the drawing, the size of the original object

may

be read

off directly.

MICROMETRY AND THE CAMERA LUCIDA.

4.

57

Determination of the Magnification of the Micro-

scope.

If the

image of a stage micrometer be projected

in pencil,
it

upon paper and drawn

possible,

will obviously

be

by comparing the actual

size of the

drawing
scale, to

with the

known dimensions

of the

micrometer

measure the magnification effected by the microscope.

The

size of the picture will,

however, vary with the

lucida,

disit

tances between the paper

is

and the camera

and

necessary to adopt for this distance a standard value,

generally 250

mm.
REFERENCES.

Carpenter, W. B., and Dallinger and its Revelations. London, 1901.

Gage,
S.

W. H.

The Microscope

H.

The Microscope.

Ithaca,

N. Y., 1904.

CHAPTER

V.

THE MICROSCOPY OF THE COMMON STARCHES.

I.

Origin and Nature

of

Starch.

^AU

living

things

depend for existence upon the potential energy of organic

compounds which they take

to simpler substances;

in as foods to

and break down

this constant

and

make good

drain upon

the limited stock of organic matter in the

is

world there

only one source of supply, the chlorophyll

bodies of green plants.

Here rays

of sunlight are absorbed,

and by
to

their

energy carbon dioxid and water are united

form

starch,
is

oxygen being

set free;

the kinetic energy

of the sun

thus transformed and stored up in potential

of

form.

Ultimately the whole structure

organic

life

depends on
rested

this process, as the universe, of mythology,

upon
is

the back of the fabled tortoise.

Starch

an insoluble carbohydrate substance belonga white powder made up of

milli-

ing to the cellulose group and having probably the formula

(CjHioOs)!.
It occurs as

small grains a few hundredths, or rarely a tenth, of a

meter in diameter, of a shape varying with ihe plant by

which

it

is

formed, but more or

less characteristic for

each species.

The

grains are in general built

up

of con-

centric layers, the outer one resembling cellulose, while

S8

MICROSCOPY OF THE COMMON STARCHES.

the inner layers contain

59

an increasing proportion of water.

frequently a small open space of
as the hilum.

At the center there

is

much
seen

less density,

known

Starch

may be

iti

situ in the chlorophyll

bodies of
it is

many

leaves in

process of active vegetation;

but

rapidly changed to
is

sugar by the enzymes of the plant, and in that form

conveyed to the root, the tuber, the stem, or any other convenient tissue, where
it

and stored for future

rich in

may be reconverted The seeds are use.

into starch

particularly

such reserve material, and with underground roots

and stems they furnish the principal sources of starch as

a commercial product.
2.

Refining of Starch.^

The processes of starch manumechan-

facture are in general simple, involving only the

ical

separation of the grains from the tissue in which they

are

embedded.

Thus, in the production of potato-starch,

the tubers after

to

washing are ground up in a comminutor

walls as far as possible.

break the

cell
is

The milky

fluid

produced
'/tio

passed through a sieve with openings

about

of

an inch in diameter which allow the grains

pulpy

to pass while retaining a large proportion of the

tissue.
ties
still

The

starch

is

then separated from the impuri-

present, either
it

by

settling, in

tanks or in long

troughs through which

gal machines; finally

flows very slowly, or

by

centrifu-

it is

dried

and barrelled

for market.
fifth of

In potatoes the starch makes up perhaps a

total

the

weight and four-fifths of the dry substance; but in

it is

the various grains

found in combination with

etc.,

nitro-

genous compounds, glutens,

plicate the refining process.

which somewhat com-

Corn-starch

may

be sepa-

6o

ELEMENTS OF APPLIED MICROSCOPY.

and
is

rated with comparative ease,

the starches;

the cheapest of

all

but in wheat and rice the union between

is

carbohydrates and albuminoid matter

introduce serious
difficulties.

so close as to
it is

In the

latter case

nec-

essary to use strong alkalies;

and with wheat the same

end has generally been obtained by moistenirg and thus

swelling the grain, bruising
it

and then allowing

it

to

stand and ferment until the nitrogenous substance has

been partly rotted and disintegrated.

feature of this

The

unfortunate

method

lies in

the necessary loss of the

glutinous constituents of the grain which possess high

nutritive value;
in certain processes the fermentation
is

omitted and gluten and bran are obtained as by-products.

The
terials.

cost of the various starches

depends both on the

of the

difficulty of separation

and on the value

raw malist

Its

range

is

indicated

by the following

of

prices compiled

by Dr. H. W. Wiley

for the year 1899:

MEAN WHOLESALE PRICE OF VARIOUS STARCHES.

Cents per pound.

Corn
Sago flour
Potato

.46-1.61

3 73-3 98

Tapioca

flour

4 21-4 57 4 63-4 96
S 7

Wheat
Rice

00-9 00
50-9 00

3.

Commercial Uses of Starch.

Wheat-starch
but
it

was

well

known
first

to the ancient
it

Greeks and Egyptians, having

said, in Chios,

been

produced,

is

was not

MICROSCOPY OF THE COMMON ST^iRCHES.

until the seventeenth

6i

century that the potato was used as

is

a source of carbohydrate; and corn-starch

still

of course a
far the

more recent product.

These three are by

most important of the starches.

As might be

inferred
is

from the scale of prices given above, corn-starch

for

used

most purposes, the amount produced in the United neighborhood of

States in 1903 being probably in the

75,000 tons.

Potato-starch ranks next with a produc-

tion of 16,000 tons in 1903,

and the amount

of wheat-

starch

was perhaps 10,000

tons.

The purposes for which this great supply is may be grouped roughly under four, heads:

intended
starch
is

used for a food material, for stiffening and sizing, as a

powder, or as a raw material for the manufacture of

other
the

substances.

Mixed with nitrogenous

supreme importance
is

bodies,

in

form of

flour, its

of course as a

nutritive substance;

and even the

no

purified product, in the

case of corn-starch, forms

insignificant contribution to

our dietary.

The

sago and tapioca flours are mainly

used for food, and the various arrowroots contain starches

recommended as

particularly desirable for invalids.

The

principal commercial importance of refined starch,

however, comes from the fact that

hot water
its

when
burst,

in contact with

grains swell

up and

forming a thick

adhesive paste.

This

may be

best brought about

by

mixing starch with cold water and slowly pouring the

thick milky fluid into boiling water
agitated

which

is

meanwhile

by constant

stirring.

The

paste thus formed

imparts to textile materials a high degree of lustre

that stiffness

and

from which the name of starch (German,

62

ELEMENTS OF y4PPUED MICROSCOPY.

Its adhesiveness Its stiffening

Starke) was originally derived.

makes
power

0t

desirable for bookbinder's paste.

it

gives

supreme importance

in the laundry;

for society

has not yet outgrown what the Puritan divine described

as

"a

certaine kinde of liquide matter

which they

call

starch, wherein the devill hath willed them to wash and

drie their rufHes,

stiffe

which when they be

dry, will then stand

and

inflexible

about their necks."

The

quality of
in

forming a

size gives starch

an increasing importance

cotton- and paper-mills, print-works

and

bleacheries.

As a powder,
preparations,
in

starch

is

used in

many
and

pharmaceutical
other products

baking-powders

where some

finely divided neutral substance is desirable,

and

for

powdering the forms

starch

in printing-houses. in the dry condition to

Finally,

when heated

i5o-2oo F. yields a soluble dextrin, sometimes called British

gum, extensively used as a

treated with
is

substitute for

gum

arable.

When
starch

weak

acids or with certain enzymes

converted into dextrins and sugars and

may

thus

serve as the

raw material

for the

manufacture of glucose,

maltose,
4.

and ultimately

of alcohol.

Microscopical Examination of Starch.

Since

the

chemical composition of the various starches

the

is identical,

microscope

offers

the

only satisfactory means of

studying them.

Fortunately the appearance of starchis

grains from the various groups of plants

teristic;

quite charac-

and one familiar with the commoner forms can

easily detect the sophistication of other substances, like

mustard, by means of starch,

bodies

the presence of foreign

(minerals or seed-hulls) in starch, or the adul-

MICROSCOPY OF THE COMMON STARCHES.

teration
variety.

63

of

one kind of starch with another cheaper

Being
rably

finely

powdered, starch

is

in a condition admi-

adapted for microscopic examination, and needs

only to be

mounted

in

some medium which

contrast.
If

will set off its

characteristics

by proper

examined dry, the

edges of the grains appear so black as to obscure the

view,

and most

solid

mounting media have on the other

hand so high an index of refraction that except with

polarized light the starch

becomes too

faint.

Water with
and
if

ordinary illumination gives a good picture;

the

hilum

is

to
like

be particularly studied, a
clove-oil

medium
staining

of higher

density

detection of starch

may may be

be useful.
aided by

Sometimes the
it

with a

dilute solution of iodine,

which produces the blue-black

color of the iodo-starch reaction.

5.

Potato-starch.

Potato-starch
in

is

produced in con-

siderable

amount both
States,

New
it

England and in certain

Western
sizing of

and

is

used mainly in print-works for the

is

warp yarn before

woven.

Other starches

make a more even and permanent

for
this

paste, as a rule, but

particular

purpose

potato-starch

seems

best

adapted.

As viewed with a hand-lens potato-starch may be

once distinguished from most Qther varieties
that a

at

by the

fact

mass of

it

appears to be studded with glittering

wheat-starch
is

points, while corn-starch or

of a dull
is

dead

white.

Under the microscope potato-starch

of large grains, .05-. 12

seen to be

made up

mm.

long, of a flattened
outline.

ovoid shape, with a smooth

and regular

As

in

64

ELEMENTS OF APPLIED MICROSCOPY.

there
is

all Starches,

a considerable variation both in shape

and

size;

many

grains are ellipsoidal,

som;

three-cor-

nered, and the smaller ones almost spherical.

Near one
dot,

end of the grain the llilum appears as a well-marked

surrounded by rather faint concentric
ellipses

which form

an important

characteristic

of

potato-starch

and

are

known
more

as the oyster-shell markings.

Sometimes two or
of
in

hila

appear in a single grain, and often groups

to

two or three grains seem

Fig. 28 (5).
6.

grow

together, as

shown

Wheat-starch.

Wheat-starch
its cdst, is
is

makes a very even

paste and, in spite of

used in

many
It is

processes

where especially

fine

work

necessary.
is

mixed with

the colors in printing cloth

ing, dyeing,

and

utilized in the bleach-'

It is also largely

and

finishing processes.

used in paper-mills.
Microscopically,
the

grains

of

wheat-starch

usually

appear as somewhat irregular

circles,

but when tipped up

on edge

their true

shape

is

seen to be lenticular.

In

size
it is

they vary from very minute points up to .04 mm., and

noticeable that the grains are mainly of two sizes, quite

large
(Fig.

and quite
28
(i)).

small, intermediate grades

being rarer
out

The hilum can sometimes be made

in the largest grains;

tric

under the same conditions concen-

circles

may

indicate the natural layers of the grain,

though as a rule neither hilum nor markings are apparent.

7.

Corn-starch.

Corn-starch,

as

we have

seen,

is

in

America by

far the

most important of starches, being used

in the kitchen, the laundry,

and the

mill for diverse pur-

poses, as well as for the

manufacture of dextrins and


MICROSCOPY OF THE COMMON STARCHES.
sugars.
It

65

may

often be found as an adulterant in the

more

costly starches,

and

in spices

and other

foods.
size.

Its grain is easily recognized,

being of

medium

".^"o:

tS
o

Fig. 28.

^The
1. 2.

Commoner Starches.
240 diameters.

(Redrawn, after Scliimper.)

4.
5. 6.

Wheat-starch.
Corn-starch. Rice-starch.

3.

Tapioca-starch. Potato-starch. Bean-starch.

.0I-.02

mm., and

of a characteristic polyhedral form,

it

its

angularity at once distinguishing

from the starches

of

wheat and potato.

Since

it is

not flattened like the

latter,

66

ELEMENTS OF APPLIED MICROSCOPY.

it

but approximately isodiametric,

in air-

shows very black edges

and water-mounts.
is

Concentric layers are absent;

but the hilum

sharply marked,

showing in water-

mount
effect

the

form

of a cross with cracks radiating out

toward the periphery of the grain.

disappears and the hilum
is

In denser media

this

seen to be round (Fig.

28

(2)).
8.

Rice-starch.

Rice-starch
It
is

resembles

that

of

corn

in a general

way, being polyhedral in shape with

angles.

distinct

facets

and

very

much

smaller,

however,

having an average diameter of about .004-.008 mm., and

the grains are rather regular in size.
istic is

Another character-

found in the

fact that the grains

commonly occur

aggregated in masses.
(Fig. 28 (3)).

As a
is

rule no hilum can be seen

This starch

used to some extent as an

of various powders.

|adulterant,
9.

and as a constituent
of the

The Starches

Pea and Bean.

The pea and

acin

bean, as well as certain other plants of the order Leguminosae, have starches of a very characteristic type; and

though not of special importance in themselves, an

quaintance with their appearance
is

an important aid

the detection of adulteration with the ground-seeds

these species.

^f

Bean-starch, which

may

be taken as an
elliptical or kid-

example of

this group,

has grains of an
.02-.06

ney shape with a

the form of a
slit

size of

mm.

running the long

The hilum has way of the grain,

while distinct concentric layers

may

be made, out in a

good
10.

light (Fig. 28 (6)).

The Arrowroots.

^Arrowroot-starches

are

exteii-

sively

used as foods for invalids and for certain other

MICKOSCOPY OF THE COMMON STARCHES.

special purposes,

67

and are rather

liable to sophistication.

The

characters of the group vary widely, and

we can

only

consider one

common
its

example.

Bermuda arrowroot has

and the posses-

a starch which in

flattened ovoid shape

sion of oyster-shell

markings resembles that of the potato.

The

grains are however smaller, .02-.03

mm., and
is

the

hilum, usually at the larger end of the grain,

typically

elongated to the form of a

slit.

The

smaller grains are

not rounded as in potato- starch, but have the same shape

as the larger ones.
II.

Sago and the Cassavas.

^Tapioca

is

derived from

the root of the bitter or poisonous cassava, the hydro-

cyanic acid which

it

contains being driven off in the pro-

cess of manufacture.

The

sweet cassava, in which this

substance

is

not present, yields a similar starch which

has recently been introduced into the trade for certain

sizing processes to

which

it

is

supposed to be specially
ciris

adapted.
cular

Tapioca starch-grains are very smoothly

and often show a truncated end.

and sHt-Hke.

The hilum

central

The size is

.01-.02

mm.

(Fig. 28 (4)).

The

grains of sago-starch have a similar hilum, and

occasionally

show truncated

ends.

They

are larger than

irregu-

those of tapioca (.02-.05 mm.),

lar,

and phenomenally

the roughly ellipsoidal form being generally distorted.

REFERENCES.
Galt, H.

The Microscopy
London, 1900.

of

the

more commonly occurring

1904.

Starches.

Leach, A. E. Maurizio, a.

Food Inspection and Analysis. New Yorli, BerHn, 1903. Getreide, Mehl und Brot.

ScHiMPER, A. F.

W.

Anleitung zur mikroskopischen Untersuch-

68

ELEMENTS Oh APPLIED MICROSCOPY.

ung der vegetabilischen Nahrungs- und Genussmittel.
1900. Jena,

VoGL, A.

E.

Die

wichtigsten

vegetabilischen

Nahrungs-

und

Genussmittel.

Wiley, H. W.
Cassava.
istry.

Potatoes and Department of Agriculture, Division of ChemBulletin No. 58. Washington, 1900.

Wien u. Leipzig, 1899. The Manufacture of Starch from

S.

U.

CHAPTER

VI.

FOODS AND DRUGS AND THEIR ADULTERANTS.

I.

Microscopical Examination of Foods and Drugs.

of

In the examination of foods and drugs for the detection of

adulterants the microscope
in
is

of very great value;

and

many

cases

it

furnishes the only satisfactory

says, in his treatise

method

analysis.

Mr. A. E. Leach

on "Food

Inspection and Analysis":

"The chemical
and the

constants of
spices

many

of the adulterants of coffee

do not

always differ sufficiently from those of the pure foods in

which they appear to be distinguished therefrom with

accuracy and confidence by a chemical analysis alone.

On

the other hand, one

who

is

familiar with the appear-

ance under the microscope of the pure foods, and of the

starches
ants,

and various ground substances used as

adulter-

can with certainty identify very minute quantities

of these materials,

when

present, with the

same ease that

one can recognize megascopically the most familiar objects

about him."
starches,

The

already treated in Chapter V, furnish

perhaps the best examples of a case in which foreign

materials

present

may be

readily
of

detected under the

microscope.

The

identification

such substances as
69

70
coffee

ELEMENTS OF APPLIED MICROSCOPY.

and mustard, containing a number
of

complex

tissues, is

more

difficult,

but a careful comparison with

samples of known purity insures reasonable certainty.

The

technique

of

the

microscopical examination
is

is

very simple.
fine (so as to

The

substance to be observed

ground up

pass through a 6o-mesh sieve), further comfingers,

minuted between the

mounted

in

water,

and

examined

directly.

It is

sometimes advisable to rub the

powder

to

still

finer condition

by manipulating

the

cover-glass

and

slide

between the thumb and

finger.

The

process should not be carried too far, because in

tissues

which are broken up into very

fine

fragments the
Bet-*

characteristic structures are often indistinguishable.

ter

views of the structure of opaque objects

mayb6

obtained by clearing or rendering them transparent by

the
action
of

strong alkaUes, caustic soda, or chloral

this
is

hydrate.
necessary.

In practice, however,

not generally

The
and

student should be careful that whatever comes in

is

contact with the specimen to be examined

free

quite clean

from the contamination

of previous specimens;

Several samples from different portions of the material

should be examined in order to gain an idea of

its

aver'

age composition.
tissue

Since the recognition of fragments of

depends largely upon the personal element, certain

points catching the eye of each observer, the comparison

with standard pure samples should never be dispensed

with.
2.

General

Nature -of

Food Adulterants.

is

^A

very

thorough and systematic examination of foods

carried

FOODS AND DRUGS AND THEIR ADULTERANTS.

?!

on by the State Board of Health of Massachusetts;

and

the

annual
as
to

reports of

that

board furnish valuof

able data

the

actual

condition

commercial
the

substances.

Spices,

coffee,

and

cocoa

are

most

important
sis is

foods

for

which the microscopical analytea, tobacco, all-

found available; while chocolate,

cassia,

spice,

pepper, cayenne, paprika, cloves, ginger,

mustard, nutmeg, vanilla, cardamom, and numerous other

materials

may be examined

to advantage.

Cocoa

fre-

quently contains foreign

fee
is

cereals, starch, or sugar;

cof-

adulterated with pea-hulls, peas, chicory, wheat,

and charcoal.

On

the other hand, in

some

of the foods

advertised as substitutes for coffee a considerable admixture of coffee

may be

found.

Tea, cloves, pepper, and

mustard are most frequently adulterated with refuse portions

of the plant in question

tea-stems,

clove-stems,

pepper-sheUs,
sionally very

and

mustard-hulls

respectively.

Occa-

bad samples

of cloves occur with a large

proportion of wheat, turmeric, and charcoal; of mustard,

nine- tenths
olive-stones.
It

wheat and turmeric;

of pepper, one-third

would be unprofitable, even

if it

were possible, for

the student to attempt to cover the whole field of microscopical analysis, since the detailed information involved

can best be acquired in practice when

it is

needed.

We
their

shall therefore take

up only

three of the

most im-

portant substances,

coffee,

mustard, and pepper, with

commonest

adulterants, as types of the rest,

and

as

illustrating the sort of characteristics

by which vegetable

food substances are identified.


72

ELEMENTS OF APPLIED MlLKU:>LUrr.

3.

The Microscopic .Structure

is

of Coffee.The coffee-

bean

the seed of Coffea arabica, a small tropical tree of

the family Rubiaceae, two of the semi-ellipsoidal beans

lying base to base in each of
its berries.

The beans

are

brought into the market

original

generally roasted, either in their

is

form or ground, and although adulteration

in the

most easy

second condition,

artificially

modelled

beans of foreign material are not imknown.

The

true

coffee-bean

is

made

of thick-walled

cells,

approximately isodiametric, and packed with

finely

Fig. 29.

^Microscopic STRUcrtrRE of Coffee. 240 diameters.

(After

Schimper.)

granular
cells

material

containing

minute

oil-drops.

The

of the inner part are very characteristic, showing

knotty thickenings of their walls, as indicated at

Fig. 29.

in

More

peripheral cells (C) are smaller and lack

these swellings, while at the extreme outside of the bean

is

the so-called silverskin, a .thin glistening layer contain-

ing pecuhar fusiform cells with wide walls pierced by

FOODS Ano DRUGS AND THEIR ADULTERANTS.

channels {A, Fig. 29).
rule,

73

The

first

two

tissues appear, as a
cell

as large

opaque dark-brown masses, with the

at the edge.

structure

showing only

The

detection

under the microscope of other

mentioned
at

cell

structures than' those

once serves to indi-

cate the presence of adulterants.

Leach has pointed out

sometimes reveals

that even a naked-eye

examination

foreign substances.

"The

chicory grains are apparent

from their dark and somewhat

gummy
and
of

appearance, and

can usually be recognized by crushing them between the

teeth.

Their

soft consistency

bitter taste are very

distinctive.

The

dull
is

surface
in

the

outside

of

the

crushed coffee-grains

marked

contrast to the pol-

ished appearance of the surface of the broken peas or

beans, often to be found as adulterants, while fragments

of

broken cereal grains are readily distinguished from

with a low-power magnifier, though perhaps not

coffee

easily identified
4.

by the eye

alone."
of
Coffee.

Chicory and other Adulterants

is

The

roasted root of Cichorium Intybus

one of the substances

coffee.

most commonly found mixed with

is at

Its presence

once betrayed by the appearance of masses of tissue

of elongated cells of the type

made up

common

to the

stems of the higher plants.

The
woody

outer layers are thintissue

fine

waUed and

delicate, while the

cells

proper contains
scar-like
cross-

large fusiform

marked with

hatching, the ladder-cells of the fibro-vascular bundles.

Occasionally, too, particles will be found whicTi

show

the

very peculiar branching, tubular structures of a

homo-

geneous dark color, which are

known

as the milk-tubes.

74

ELEMENTS OF /IPPLIED MICROSCOPY.

general predominance of elongated
cells,

The

the presence

of ladder-cells

and

milk-tubes, the absence of opaque

masses of dark-brown tissue and of granular cell-contents

make

the picture very different

from that obtained

in the

case of coffee.

Various other roasted roots,

fruits

radishes,

carrots,

etc.,

such as

figs

and pears, with grains

of diverse sorts

are sometimes used as coffee substitutes.

setts the following

In Massachu-

have been found

Roasted peas, beans,

wheat, rye, oats, chicory, brown bread, pilot-bread, charcoal, red slate, bark,

and dried

pellets, the latter consist-

ing of ground peas, pea-hulls, and cereals, held together

with molasses.
cells,

In each case the presence of pecuhar

other than the simple ones characteristic of the

coffee-bean, will reveal the sophistication.

The

groundfor

up

seeds of the Leguminosae, peas

and

pea-hulls,

example, are quite commonly present, and

tected

may

be de-

by the

starch-grains described on p. 66, and by

the outer layers of the hull, which from their parallel

structure are
5.

known

as the palisade

cells.

The Microscopic Structure

of Mustard.
is

^Mustard,
in

as

it

usually comes to the market,

a mixture of the

ground seeds of two

Brassica
(Sinapis)
is

closely related herbaceous plants,

alba

and nigra.
if

The

structure,

both cases,

very similar, and

the entire seed be ground

up and examined,
rent.

three distinct cell types will be appaflat trans-

On the

outside are several layers of large

cells,

parent thin-walled
onal,

some rounded and some

faint
d,

polyg-

showing in

glycerin
(c

concentric

markings
epider-

about a median area

and

Fig. 30).

These


FOODS AND DRUGS AND THEIR ADULTERANTS.
mal
layers
lies

75

are

difficult

to

them
layer

a characteristic

make out tissue known

clearly.

Within

as the

columnar
under the

made up

of prismatic cells which, as seen

microscope, are small and polygonal with heavy walls

and small central lumens, giving the whole a dotted appearance.

lowish,

In white mustard {B. alba)

this tissue is yelit

while in black mustard {B. nigra)

is

dark

brown, determining in each case the general color of the

Fig. 30.

^Microscopic Structitre of MtrsTAKD. 240 diameters.

(After Schimper.)

seed

(h,

Fig.

30).

Finally,

the interior
cells,

is

a tissue of

medium-sized, fairly thick-walled

fine,
{a,

packed with a
.

gray-green, granular material containing oil-drops

Fig. 30).

6.

Adxilterants of Mustard.

^In

a good table-mustard
tissue present

the

amounts of epidermal and columnar

are very small, since the hulls are largely sifted out; the

76

ELEMENTS OF APPLIED MICROSCOPY.

cellit

bulk of the sample should be made up of the inner

contents.
is is

If

an excess of the outer layers be present,

evident that mustard-hulls have been added; and this

one of the commonest adulterations to which this subis

stance

subjected.

Wheat- and

rice-starch are

also

found not uncommonly, and furnish conclusive evidence

of
sophistication.

Yellow aniline dyes are sometimes

used to give the proper color to such adulterated samples.

Turmeric, which
longa,
tard.
is

is

the

ground rhizome of Curcuma

also well

adapted for the adulteration of mus-

It

appears under the microscope in small amor-

phous, intensely yellow, pasty masses, which stain blue

with iodin, being

7.

made up

largely of curcuma-starch.

The Microscopic Structure

of Pepper.

Pepper

is

the pulverized seed of Piper nigrum, a shrub cultivated

mainly in the East Indian

about

islands.

The
is

dried seed

is

5 millimeters in diameter

and

covered with a
grain,
is

brownish

hull.

If this hull is

ground up with the

pepper.

we have the ordinary black made by macerating the fruit

detaching the
hulls,

White pepper

in water before drying,

and

by

friction.

In ground black pepper a number of tissues

may

be

made

out,

since

the many-layered hulls are relatively

central

thick as
portion.

compared with the more homogeneous

The

latter

forms the bulk of the preparation,

It consists of

and

is

quite characteristic in appearance.

irregularly angular whitish masses, seen to be

made up
these

of polygonal cells

packed
Fig.
31).

full

of very

minute starch-

grains

{s

and

ss,

With a high power

and

grains themselves appear polygonal

closely aggre-


FOODS AND DRUGS AND THEIR ADULTERANTS.
gated into masses, while their size
is

77

very small, averaging

about .003

mm.

Besides these starch masses at least

two

characteristics

kinds of

cells

from the hull

may
so

appear in black pepper.

The outermost

layer furnishes
stone-cells

dark-brown fragments made up of the

called, polygonal,

thick-walled cells with a small dark

lumen and

striations radiating out

from

it

(a.

Fig. 31).

Fig. 31.

^Microscopic Structure of Pepper. 240 diameters.

(After

Schimper.)

The

cells of the

inner part of the hull occur in lighter,

yellowish-brown masses, and have a somewhat similar

structure except that their
their

lumen
31.)

is

much

larger

and

waUs thinner
tissue

(6,

Fig.

Smaller fragments of

parenchyma

may

be seen, with irregular, thin-

waUed
of oil

cells

and occasional large spaces where deposits

fruit (c, Fig. 31).

were present in the

White pepper made up simply of the central part of

the seed contains practically nothing but the irregular,
silvery

masses of pepper-starch with occasional small

fragments of the outer tissues.

78
8.

ELEMENTS OF

/IPPLIED MICROSCOPY.

Adulterants of Pepper.
adulterated
of

Pepper
spices,

is

one of the most

commonly

and may be found

mixed with a wide variety

of mustard, the

of substances.

As

in the case

ground hulls of the seed

itself

furnish a

frequent sophistication.
the United States
is

The most
it

general. adulterant in
is

perhaps buckwheat-starch, which

not easy to detect since

closely resembles that of the

pepper in shape and occurrence.

are,

The

individual grains

however, about twice the size of the pepper-starch,

as a rule, are also larger.
is

and the masses,

In Germany

dried and ground bread

often used, as well as the bark

of trees, bran, sawdust, pulverized nut-shells, of mustard, rape, peanut, linseed, or almonds.

and

hulls

In France

and
form

in this country

ground oHve-stones are a .common

adulterant.

They

are

made up

principally of large fusi-

stone-cells

which resemble those of the pepper

except in their size and the fact that they are practically
colorless.

The

occurrence of any substance other than the

normal
starches
tic of

tissues of the pepper,

and

in particular of foreign
characteris-

and the elongated

cells

and tracheids

plant-stems, will suggest the presence of

some

adul-

terant.

In

all

these instances only careful comparison with a

series of

known pure

substances will enable the analyst

is

to determine certainly which of these foreign materials

present.

FOODS ^ND DRUGS AND THEIR ADULTERANTS.

79

REFERENCES.
Greenish, H. A.
Drugs.

The

Microscopical Examination of Foods and

Philadelphia, 1903.

KoNiG,

J.

Die

menschlichen

NahrungsAnalysis.

und

Genussmittel-

Berlin, 1904.

Leach, A. E. Leach, A. E.
tion.

Food Inspection and

New

York, 1904.

Microscopical Examination of Foods for Adultera-

Thirty-second Annual Report of the State Board of

Health of Massachusetts for 1900.

MoELLER,

J.

Mikroskopie der Nahrungs- und Genussmittel aus

Berlin, 1886.

dem

Pflanzenreiche.

ScHiMPER, A. F.

W.

Anleitung zur mikroskopischen UntersuchJena,

ung der vegetabiUschen Nahrungs- und Genussmittel.

1900.

ViLLiERS, A., et Collin, E.

Traite des alterations et falsifications

Paris, 1900.

des substances alimentaires.

TscHlRCH,
VoGL, A.

A., u.

Oesterle, O.

Anatomischer Atlas der PharmaLeipzig, 1900.

kognosie und Nahrungsmittelkunde. E.

Die wichtigsten vegetabiUschen Nahrungs- und

Berlin, 1899.

Genussmittel.

CHAPTER

VII.

THE EXAMINATION OF TEXTILE

I.

FIBRES.

The Kinds
the

of Textile

Fibres.

^The
is

word

fibre

is

derived from
or filament."

Latin

jihra,

and

signifies

"a thread

The most

primitive application of natural

fibrous materials to textile purposes

probably in the

use of grasses and osiers for weaving.

period,

At a very

early

however, primitive

man

learned to manipulate

fibres of a closer texture in the

cloth.

manufacture of coarse

The
races

long, fine hairs of plants

and animals among

attention;

many

must early have attracted

In Egypt

thus
be-

the application of cotton

and wool dates back from

yond recorded
bast-layer

history.

the use of the fibrous

from the stem

of the flax-plant appears to have

antedated even that of cotton.

According

to.

Chinese

tradition, the application of the secretion of the

silkworm

to textile purposes

was made by the

hemp,

first

empress of the
the prin-

nation.

To-day, four classes of fibres

flax,

cotton,
the

cipal vegetable hair;

all

jute, ramie,

and

sisal,

typical bast-fibres;

wool and a few

less well

known
of

animal hairs;

and

silk

still

make up

textiles

most importance.

The

leaves of certain plants furnish

filaments essentially similar to the bast-fibres mentioned

80

EXAMINATION OF TEXTILE
above.

FIBRES.

8i

The

leathery covering of the cocoanut suppHes

a fibre used for matting, etc.

Asbestos illustrates the

possibility of using a mineral substance for textile pur-

poses.

Finally,

various

artificial

fibres are

made from

metals and from cellulose derivatives.

In the identification of
of

textile fibres the

microscope

is

prime importance.

Animal
of

fibres in general

may
by

be
the

distinguished
fact that

from those

vegetable

origin

both wool and

silk are soluble in

5%

caustic

soda, while the vegetable fibres, are not thus affected.

of

made up
of

of cellulose,"

Various color reactions are also

individual fibres,

value.

For the determination

is

however, the microscope

most

satisfactory, since
its

even

of the closely related bast-fibres each has

tic

characteris-

appearance.

Furthermore, the quality of different

is

samples of the same kind of fibre

size,

evidenced by the
of the filaments.

twisting,
effect of

and external structure

The

chemical reagents and the tensile strength

of individual fibres

under various conditions

is

studied

with the aid of the microscope in modern

tories.
2.

textile labora-

The

Cotton-fibre.

The
down

cotton-fibre

is

the vege-

table hair borne

on the seed of plants

of the

genus Gos-

sypium, serving like the

of the thistle for the distri-

bution of the seeds by wind.

the

These plants belong

to

Mallow family;
to

some are herbaceous and others

grow

be bushes twenty feet in height.

As

the seedof

capsule opens a rich white boll bursts out,

the cottonseed with the fibres at
its

made up

free end.

The

cotton-plant grows well

between 45 north and

82

ELEMENTS OF APPLIED MICROSCOPY.

35 south latitude, given proper soil and a uniform supply

of

moisture.

The United

States,

India,

Egypt,

China, and Brazil are the most important cotton-pro-

ducing nations in the order named; though

countries devote large areas to
its

many

other

cultivation.

The most important

ton for the market
fibre
is

process in the preparation of cotginning, or the separation of the

from the seed-cotton, which as picked contains

its

two-thirds of

weight in seeds.

The modem

process,

evolved from that which Eli Whitney was largely instru-

mental in developing in 1794, consists in exposing the

cotton to the action of a series of fine-toothed circular

saws which tear

off the fibre

and carry

it

away through

the grid in which they revolve.

The

fibre,

as thus obtained,

is

a hollow ribbon

(Fig.

32) spirally twisted at frequent intervals with the edges of

the ribbon so sharply jnarked off from the central canal

that they appear like swollen cords.
fibre
is

The

section of the^
this

not,

however, dumbbell-shaped, as

might,

suggest, but elliptical or

somewhat

crescentic, the

lumen
is

of the canal following the outer contour.

The

canal

narrow in American and Egyptian cotton,

that

and. broader in

from India.
it

By

following a single fibre carefully

it

along,

will

be noticed that

tapers to a somewhat
is

blunt point at one end and at the other extremity

broken

off

sharply where

it

was attached

to the seed.
40,

The
mm.,

length of the cotton fibre varies from 20 to

its

diameter, from 10 to 20
is

//.

The

cotton-fibre

mainly

cellulose,

covered with a

fine

cuticula of different composition.

When

treated with a


EXAMlN/ITlON OF TEXTILE FIBRES.
solution of hydrate of copper in
dissolves

83
cellulose

ammonia, the
into bubbles

and swells up the cuticula

between

the twists of the fibre, while at the twists the cuticula contracts, giving the characteristic

beaded appearance shown

in Fig. 31,

e.

Much

of the cotton in textile goods has been subjected

Fig. 32.

^The Cotton-fibre.

(After Hassack.)

200 diameters.

to the process of mercerization, or

immersion of the cloth

soda solution.

in a stretched condition in strong caustic

Cloth thus treated

is

stronger, takes dyes better,

and has

closer, firmer texture

and a

fine glossy

appearance.

Under
central

the microscope the individual fibres appear as crinkly,

irregular cylinders instead of flat ribbons

and the

canal

is

much

shrunken.

Sometimes

the fibres

are sq


84

ELEMENTS OF APPLIED MICROSCOPY.

swollen that even the twists cannot easily be seen in a

water-mount; they show more

3.

clearly,

however, in
of

air.

Bast-fibres in

General. Most

the

vegetable

fibres in

commercial use are derived from the bast-layer

of the dicotyledonous flowering plants.

The bast,
more or

or, as it is called

by botanists, the phloem,

is

fibrous layer lying just

less

under the bark, and since

all

it- is

developed in

the higher plants,

textile fibres

it

would

be theoretically possible to obtain

from any

one of them.

In preparing the

fibre

from those herba-

ceous plants actually in commercial use, the stems are

first

retted or allowed to ferment

under water so that the

gummy

substances which hold the tissues together

may

be dissolved.

Next they are scutched or exposed

to the

action of beaters which break

up

the outer and inner

friable tissues, leaviftg the elongated bast-cells adhering

together in threads or bundles.

find
it

The

bast-fibre, as.

we

in

commerce,

is

thus

made up

of a group of

cells,

not of a single

cell like the cotton-fibre.

When
bast-cell

further broken

up under the microscope,

the

appears as a more or less elongated spindle

with a central canal where the living protoplasm once

lay, the cell wall of cellulose

only being

left.

Both ends

are symmetrically pointed.

section,

In

size,

proportions, crossof various

canal,

and markings the

distinguished.

bast-fibres

plants

may be

We

shall consider briefly

the six fibres which have most commercial importance

flax,

hemp, Manila hemp,

4. Flax.

jute, ramie,

and

sisal.

^Flax is
tall

obtained from the stem of

Linum

usitatissimum, a

herb widely cultivated in the central


EXAMINATION OF TEXTILE
regions of

FlBf^ lES.

Si

Europe and America.

The

seed of the plant

furnishes linseed-oil,
into linen.

and the bast

yields flax to be

spun
a

The
cells,

flax-fibre like others of its

class, is

bundle of
ends

each a hollow cylinder tapering at both

Its
cells

(Fig.

33).

are

distinguished

by

their

d
Fig. 33.

^The Flax-pibm;.

(After Hassack.)

200 diameters.

large size (25 to

30 nun. long by

.02

mm.

in diameter),

and by the

fact that they are swollen or knotted at fre-

quent intervals.

The

central canal

is

narrow and nearly

circular in section, while the cell itseK is

somewhat

flat-

tened.

Thus when a number

some
will

of cells are

examined under

the microscope

appear

much broader than

is

others, while the canal in all of

them

narrow.

At the

end the

flax cell tapers gradually to

a very sharp point,

tip.

and the canal disappears

at

some distance from the


86
5.

ELEMENTS OF APPLIED MICROSCOPY.

Hemp.

The
is

true hemp-fibre, to be distinguished

Sisal

from Manila hemp and

sidered later,

hemp, which

will

be con-

derived from Cannabis sativa, an herb

as the flax-plant,

with

much

the

same range

grown most
a strong

extensively, perhaps, in Russia

fibre,

and

Italy.
is

It is

though

less pliable

than

flax,

and

used for cordage,

sail-making, and the manufacture of certain other textiles.

The Cannabis

plant also yields the drug hashish.

Fig. 34.

The

Hemp-fibre.

(After Hassack.)

200 diameters.

The

hemp-fibre

cell

very closely resembles that of

flax

in its microscopic appearance, being of the

same

size

and

general shape
It
..the

and showing the same knots and

cell, is elliptical
its

swellings.
First,

may

be distinguished by two characteristics.

canal, like the

cell

in cross-section, so that
distin-

while the

on

narrow surface cannot be


EXAMINATION OF TEXTILE
guished from
flax, its

FIBRES.

87

broad 'diameter shows the broad

Second, the ends of the
cell are

canal quite distinctly.

comparatively blunt, and the canal runs up to the very

tip (Fig. 34).
6.

Jute.

^The

jute-plant, Corchorus capsularis,

is

an

annual, native to the East Indies, and furnishes a long

smooth

fibre which,

however, quickly softens and breaks

Fig. 33.

The

Jute-fibre.

(After Hassack.)

200 diameters.

when

wetted.

It is

used for making rope, coarse twine,

tex-

and gunny-bags, and for the backing of various other

tile

materials.

The cells of which the much shorter than those

about
2

long jute-fibre
of flax

is

made up
is

are

and hemp, being only

width
nearly the
field of

mm.

in length, while their

fibres.

same as that of the other

the microscope

In any given

numerous ends

of cells will

be apparent,


88

ELEMENTS OF APPLIED MICROSCOPY.

while with the long cells of

are hard to find.

Linum and Cannabis

the tips

The most marked

ever, its canal,

peculiarity of the jute cell

is,

how-

which varies markedly in diameter,

at one

point occupying a large part of the cell ing to an almost invisible line (Fig. 35).
7.

and then

shrink-

Ramie.

Ramie or China grass

Islands.

is

the product of a

low shrub, Boshmeria nivea, indigenous to China, Japan,

and the Philippine

Its fibre is long

and

lustrous,

Fig. 36.

^Ramie.

(After Hassack.)

200 diameters.

stronger probably than any other fibre;

"it is

used for

making
cells of

sail-cloth

and

for other special purposes.

The
their their

ramie (Fig. 36) are easily distinguished by

great length, sometimes reaching 50 cm.,

flat

and by
they

ribbon-Hke

structure.

In

cross-section

are

EXAMINATION OF TEXTILE
pointed
ellipses,

FIBRES.

89

.03-.06x.01-.02 mm., with a wide canal

cell.

of the same shape as the

Under

the

microscope,
like

therefore, as

one sees the two views, ramie appears

fibre

mixture of a broad
fibre
8.

with a broad canal and a narrow

with a narrow canal.

Manila Hemp.

^Bast-fibres are not confined to the

stems of plants, but are continued upward in the fibrovascular bundles which form the veins of the leaves.

In

many

plants these leaf -fibres are of such a character

textile

as to
of

be valuable for

purposes, notably in the case

Manila hemp and

Manila hemp
is

Sisal

hemp.

obtained from the leaves of

to

Musa

texiilis,

palm native

the Philippine Islands and

is

North Borneo.
though more
for

The

fibre

extracted
is

by hand, and,
extensively used

brittle

than true hemp,

marine cordage.

The

cells

are distinguished from

those of Cannabis

by

their lesser length (about 6

mm.)
canal

and by

their smoothness, the knots,

though present, being

much
is

less

marked than

in the hemp-fibres.

The

wide and easily discerned.

9.

Sisal

Hemp.

Sisal

hemp

is

obtained

from

the

leaves of a cactus,

Agave

rigida, extensively cultivated in

It is

Mexico, Central America, and the West Indies.

long coarse white fibre used for cordage and the
facture of rough sackcloth,

manu-

hammocks,
all

etc.

The
by

cells

are distinguished from

other bast-fibres
are 1.5-4

their spioothness
/z

and

regularity.

They

mm.

long and 20-32

wide, approximately cylindrical, with a

wide and well-marked canal, and tapering to a fine point

at the end.

As

in other cases,

comparison with material

9
of

ELEMENTS OF y4PPLIED MICROSCOPY.

known composition

furnishes the only sure criterion

for identification.

^Animal hairs are 10. Wool and Other Animal Hairs. much more complex structures than those of plants, being
not only multicellular, but composed of several distinct
layers of cells.

In the growth of a hair the epidermis or

is first

outer tissue of the skin

folded in to form a minute

the base of this
follicle

pit called the hair- follicle.

From

the hair grows

out, its cells dividing off

from the

epider-

mal

tissue below.

The

hair itself
is

is

made up

of three

distinct zones.

In the center

an

axis of irregularly

rounded
this,

cells

known

as the medullary layer.

of the hair,
is

Outside
the corcells;

and forming the main portion

which
consists
of

tex,

elongated

spindle-shaped

and

at the periphery is a cuticular layer of flat overlapping

scales covering the hair like the shingles

on a

roof.

In
to

the cortical cells the hair

its

is

situated the pigment

which gives

color,

and

to the disintegration of these cells

is

and the formation

of air-spaces in their place

due the

whitening of the hair in old age.

Externally the principal characteristic of
hair
is

the

animal

the presence of the scales of the cuticle

which give

the fibre the effect of being

marked with
at its

fine transverse,

anastomosing
as
fine

lines

and which

edge (Fig. 37) appear

cells

serrations.

The medullary

may

just be
stria-

made
tions.

out beneath this cuticle as fine longitudinal

The

general term wool

is

applied to hairs which are

will

spirally twisted so that

when woven they

hold

to-

gether to form a strong fabric.

In such hairs,

too, the


EXAMINATION OF TEXTILE
scales are large

FIBRES.

91

and project prominently from the

fibre, so

that they interlock with each other

and materially

in-

crease the compactness of the tissue.

varieties of

Some

thirty-two

sheep (belonging to the genus Ovis) furnish

the wool of

the Thibet or

commerce besides the llama, or alpaca goat, Cashmere goat, and the Angora goat, from

Fio. 37.

Wool-fibres.

(After Hassack.)

200 diameters.

which mohair

is

derived.
its

The

length of wool varies

from

2 to

20 cm., and

diameter from 10 to 100 ^; the'

into worsted

larger fibres are

combed and spun

and spun

yams,,
7".

the shorter are carded

into woolen yarns.

The

separation of wool from vegetable fibres under

is sufficiently

the lens

simple;

but the application of thqt

niicroscope only begins here.

The

hair from various,

animals

should

be

carefully

studied;

coinparison

qi[

92

ELEMENTS OF APPLIED MICROSCOPY.

sheep's wool with camel's hair will prove instructive,

showing the

lesser length of the latter, its

flat

freedom from

twisting, the scales lying almost

against the surface,

Different grades

and the granular spots

of

in the medulla.

wool

may

then be

studied, contrasting the high-grade

15/i

merino wools with a diameter of

and 10 or more
wool of poor

twists in every centimeter of length with a

quality having perhaps four or five times that diameter

and not more than one

two
qualities, fineness

twist in a centimeter.

These
deter-

and convolution, principally

its

mine the quality

projection of
its

of wool, although

regularity

and

the

scaly covering are also of importance.

sample of wool shoddy

may

profitably be examined;

various foreign fibres, dyed fibres, and torn and broken

fibres will

be apparent.

II. Silk.

One
The

other animal fibre of quite a different

character remains to be considered, the secretion of the

silkworm.

larvae of

many moths

spin their cocoons

out of threads long and strong enough to be used for

textile purposes,

but that of

Bombyx mori

is

cultivated

most extensively for commercial purposes in China,

Japan, India, France, and Italy.

The
of the in the
fibres

substance of which silk

is

composed

is

poured out
end

in a liquid condition

from two glands

at the anterior

worm, and when discharged

form of
solid

in the air at once sets

rods.

structureless

The

pair of

formed by the two large spinnerets are cemented

together

by an incomplete

cuticula of

somewhat
set of

differ-

ent composition produced

by another

glands and

known

as sericin.

Under

the microscope such a double


EXAMIN/tTION OF TEXTILE FIBRES.
thread at
first

93

sight appears like a

broad

fibre

with a

central canal (Fig. 38, a).

ket,

In preparing

silk for the

mar-

however, the cocoons, before unwinding, are placed

in hot

water to

kill

the chrysalis
all

and

to melt the

gummy
In
this

material which cements

process the single fibres

the threads together.

become separated and each ap-

pears only as a semi-transparent cylinder (Fig. 38, b) with

Fig. 38.

Silk.

(After Hassack.)

200 diameters.

no internal structure, and no external markings except very

fine
sets.

longitudinal cracks formed

as the fresh secretion

Flecks of the ruptured cuticula and particles of

foreign material
liquid

which come in contact with

fibres.

sometimes adhere to the

it when semiThe diameter

varies considerably
rate at

from point

to point, according to the

its

which the worm was producing

/j.

secretion.

Perhaps 10-20

would be a

fair average.

Ends, except

94

ELEMENTS OF APPLIED MICROSCOPY.

accidental breakage, are, of course,

when produced by
meters of silk
12.
is

absent, since the whole cocoon containing perhaps 500

often

made up

of a single piece.

Analysis of Fabrics.

In

the examination

of

fabric a sample should be taken 2-3 sq. cm. in size, or large

in_

enough

to include all the different yarns

employed

the pattern.

The warp and

filling

threads are then

separated into their constituents, and one of each kind

taken for analysis;

different

sometimes there

may

be a dozen
of the
is

yams

to

examine.

Under a low power

microscope

(fifty

diameters) the nature of the fibres

determined in the different yams, taken in the proportion

in

which they occur in the

fabric;

a rough quantitative

analysis

may

thus be made.

REFERENCES.
Die animalischen Faserstoffe. Leipzig, 1902. Die vegetabilischen Faserstoffe. Leipzig, 1900. Bowman, F. H. The Structure of the Wool Fibre Manchester,
Bottler.
Bottler.
1885.

Brooks, C. P.

Cotton.

New York,

1898.

Dodge.
1897.

Descriptive Catalogue of the Useful Fibre Plants of the

World.

Report No. 9 of the U.

I.

S.

Department of Agriculture

Hannan, W.
Hassack,
C.

The TextUe Fibres of Commerce. London, 1902 Wodurch unterscheiden sich die Textilfasem ?

Leipzig, 1899.

Matthews,

J.

M.

The

Textile Fibres

New

York, 1904.

ViGNON, L. La Soie. Paris, 1890. Wilkinson, F. The Study of the Cotton Plant.

New York,

1899

CHAPTER Vm.
THE MICROSCOPY OF PAPER.
I.

Paper and Paper-making.

The
A real
era.

earliest

written

records were probably

stones, leaves, bits of

made on such
clay.

natural objects as

bone or wood,

or, as in

Babylonia

and Assyria, upon blocks of

the celebrated papyrus,

paper, however,
in

was manufactured

Egypt many
thin, trans-

hundred years before the Christian

The

parent layers of tissue which surround the stem of the

papjTUs plant were separated with some sharp instrument, superposed under water and then pressed anddried.

Large quantities of

this

product were exported

from Alexandria

to all parts of

Europe and Asia, coming and

into competition with

parchment, prepared by scraping

goat.

and drying the skin

of the sheep

The modem
duced
to

process of

making paper,

as a thin layer
re^

of cellulose derived

from fibrous vegetable material

first

a pulp in water, was

discovered

by the

Chinese and introduced into Europe by the Arabs in the

eleventh century.

Cotton was the

first

material used-

for this purpose, but

an Arabic manuscript on linen pap^

is
still

bearing the date iioo

facture of paper
first

in existence.
to

The

mariii-

from wood-pulp appears

have been

suggested by the French naturalist, Reaumur, in

95

96
1 7 19,

ELEMENTS OF /IPPLIED MICROSCOPY.

who
first

derived his idea from a study of the

nests ffom
is

way

which wasps construct their

this material.

The

step in paper-making

the treatment of the

raw stock with some

which

chemical in order to

break

it

up, to

dissolve the cementing

is

gums, and to separate the

cellulose,

always the principal constituent of paper, in as

pure a form as possible.

The

crude material

is

generally

boiled with strong alkali under pressure,

in a tank

and then washed

is

from which the waste water

removed by a
acid

revolving drum.

Wood

paper

is

also

made by an

process, the stock being treated with the bisulphites of

lime and magnesia;

and mechanical wood-pulp

for a

low grade of paper

is

prepared by simple attrition against

stone surfaces without chemical action.

After treatment in the boilers and washers, the paperstock

stuff."
is

bleached, and at this stage

It is next

is

known

as "half-

passed through the beaters, in which a

it

wheel bearing knives breaks

condition.

up

into

a fine fibrous

Sizing, loading, coloring, and other auxiliary

processes are accomplished in the beaters.

fully
it

Finally, the

prepared pulp passes to the paper-machines, where

spread in a thin sheet of running water over a
belt of wire cloth.

is

moving endless

The

fibrous material

deposited in a fine and even layer passes on through

felted rollers,

which press out the

last of the

water and

compact
2.

its

texture.

The

Raw

Materials

of

Paper.

Obviously

paper

might be made from any material which can be ground

up
as

to a fine fibrous pulp;

and such bizarre substances

corn-husks,

seaweeds,

shavings,

sawdust,

cabbage-

MICROSCOPY OF PAPER.

97

Stumps, and leather- cuttings have been used experiinehtally for this

purpose.

Cotton and linen rags

of

still

fur-

nish

the

best

grade
is

paper.

The manufacture

of

Manila paper

an important industry.

Straw and

wood-pulp, however, furnish by far the largest yield of

coarse paper

and cardboard.
is

The

relative

importance of

the different materials

indicated

by the following table

from the United States Census

Paper-stock Manxtfactuiied in the U.
S.

in 1900.

Wood, cords
Wood-fibre, tons
Straw, tons

1,986,310

644,006
367,305

Old paper, tons

Rags, tons

356,193
234,514
99)3!
of

Manila, tons
3.

The Microscopic Examination

Paper.

Before
it

examining a sample of paper under the microscope,

should be torn into small bits and boiled in a one-percent

solution of caustic

soda.

The wet pulp produced

fibres is

is

washed on a

fine sieve

and broken up by shaking

paper

in water.

The
with

identification of

by no means an
be

easy task, and requires careful study and comparison

known

substances.

The

student

will

much

monograph prepared by Professor Mr. A. G. Woodman, and pubW. R. Whitney and

aided by an admirable
lished in the

Technology Quarterly for September, 1902.

The
the

authors suggest the following points to be noted in

systematic study of each specimen:
size

of cells,

shape, length, width as

compared

to length,

shape of

the ends, presence of knots or joints,

whether the major-

98

ELEMENTS OF APPLIED MICROSCOPY.

bent or curled, markings,
Polarized

ity of the cells are straight or

size

and character

of the central canal, if any.

light is
fibres.

often of great service in the identification of paper

4. Analytical

Key

to

Paper

Fibres.

^The

authors

the the

above mentioned suggest the following useful key for preliminary examination of paper. In this table

word

fibre is restricted to those cells

which are very

long in proportion to their breadth.

ANALYTICAL SCHEME.
A.
Fibres are characteristic; other characteristic forms absent.
I.

Fibres are long; greater than diameter of field (Mag.

(i)

= 60).

Fibre has

many

joints, knots, or projections, especially

light,

by polarized
(a)

Fibre

is fine;

quite smooth; joints not very notice-

able.

Paper mulberry
(6)

Manila hsmp Agave.

and promisaliva),

Fibre

is

coarse; uneven; joints are large

nent.

Linen
(c)

JuteHemp (Cannabis

Fibre shows overlapping scales.

Wool.
(2) Fibre has peculiar
net-like.

markings; dotted, circular, square,

circular or square perforations.

(a)

Markings resemble
Spruce
(fir,

Redwood
(b)

Pine Red cedar Arbor vitm Cypress.

hemlock, tamarack, balsam)

Markings net-like, "feather-stitch," Redwood Cypress Banana Ramie.

spiral.

(fruit- stem)

(3)

Fibre
(o)

is

smooth and
fibres

regular.

Many

Cotton
(b)

Banana

resemble twisted ribbons.

(stalk

and

leaf-stem),

Fibres are round, cyUndrical.

Silk

Sisai hempBark

oj cotton^stalk.

y.:c..oscoPY
II.

of paper.

99

Fibres are short; less than diameter of field (Mag. =60).

(i)

Ends Ends

of fibres frayed

Cotton rag
(2)

Mechanical wood

and

torn.

(conifers).

of fibres not torn.

Coir
B.

ElmWillow.
fibres are also present.

Characteristic forms other than

I.

Fibres are long; equal to or greater than diameter of field

(Mag. =60).
(i)

Fibres quite broad in middle; tapering to point like a

bayonet,
(a)

One

or both ends of cells

drawn

out,

sometimes

Poplar
(6)

Ends

BirchPorpor gum. CottonwoodWhitewoodBlack

Chestnut.

terminating in a

tail.

of cells cut off obliquely at blunt angle.

walnut

Holly

(2)

Fibres not varying greatly in width, ends needle-Uke.

(a)

Characteristic forms (cells) large;

covering i to J

diameter of

Bamboo
(b)

Sorghum bagasse

field.

Raffia

Tulip.
serrated,

Characteristic forms small,

cellular,

or

pointed.

Straw
II.

Esparto Sugar-cane bagasseLive Oak.

and narrow;
quite sharply

Fibres are short; less than diameter of field (Mag. =60).

(i)

Cells comparatively long

pointed at one or both ends.

Magnolia
(2) Cells

Tulip Sweet buckeye.

stubby;

prismatic,

ends

blunt

and cut

off

obliquely.
(a)

Quantities of short,

fine,

transparent material with

square ends.

Holly
(b)

Chestnut.
and
characteristic

Field fairly clear except for fibres

elements.

Pawpaw
cherry
(3)

Tree Willow.
Elm.

of

heaven

Maple Black

Cells fragmentary, broken; fibres short.

Groundwood


too

ELEMENTS OF APPLIED MICROSCOPY.

With a few of the 5. The Commoner Paper Fibres. most important materials of paper the student should
thoroughly familiarize
linen,

himself.
jute,

The
silk

fibres

of

cotton,

hemp, manila,

and

have already been

treated with

some

fulness in

Chapter VII.

Of
pine,

the woods
dis-

used in the manufacture of paper pulp there are two

tinct types.

The

Coniferae

spruce,

fir,

etc.^

ex-

hibit cells of

markedly

different structure

from the Angio-

sperms, of which poplar and birch are the most important

examples.

Finally,

straw and esparto grass are

distinguished
6.

by a

third characteristic type of tissue.

The Structure

of the
fir,

G3minospenns.

^Wood-pulp
is

made from

the spruce,

balsam, larch, or hemlock

Fig. 39.

Tracheid of a Conifer.

(After Herzberg.)

240 diameters.

mainly made up of the

group the structure

cells

from the tracheids or

In
all

fibro-

vascular bundles of the stem.

is

the trees of this

essentially the same.

The

cells
field,

are long, exceeding the diameter of the low-power

and
are

of considerable breadth

(see

Fig.

39).

The

ends

often contracted to

a rather acute point.

Sharp
in

twists are

sometimes present, due to harsh treatment

the preparation of the fibre.

of these cells, however,
is

The

characteristic feature

the presence of

numerous round


MICROSCOPY OF PAPER.
,

lOI

or oval scar-like pits arranged in regular rows over their

surface.

In the case of pine,

lattice-like areas

with ob-

long openings appear at intervals; long

ings
7.

but the presence of

cells
is

mainly of a single type and with discoid mark-

the characteristic of the Conifers in general.

of

The Structure
the

the

Angiosperms.

Pulp

made
broad

from

commoner Angiospermous

trees

shows two

distinct elements,

long narrow fibres and short

Fig, 40.

^Tracheid of Birch.

(After Herzberg.)

240 diameters.

cells

with .characteristic
in

markings.

The two

species

most

use

for

cheap grades of paper are poplar

fibres are very

and birch; the longer

having
a
central

similar in both,

canal

of

variable

width and ends

sometimes rounded and sometimes tapering to a point.

The more

thin- walled of these fibres often

show rounded

and oval pores penetrating the

wall.

Smaller fibres with


ELEMENTS OF APPLIED MICROSCOPY.
swellings at intervals along their course are also present
in birchwood.

The
are

small,

broad

cells

of

the .fibrovascular bundles

more

characteristic.

In birch they are very numer-

A
Fig. 41.

B
Cells of Straw.
(After Herzberg.)

C
240 diameters.

ous,

and are dotted with minute pores arranged

in rows

at right angles to the long axis of the cell (Fig. 40).

The
At

distribution of these pores

is

somewhat

irregular.

the ends of the cell the cross-hatched transverse walls are sometimes seen.

In poplar the

cells of this

type are

MICROSCOPY OF PAPER.
less

103
closely packed.

numerous and show larger pores more

cells is

At the ends of the

often a long, tail-like point,

is

and

the grating seen in the case of birch

8.

absent.

The Fibres

of

Straw and Esparto.

Paper
largely

made**

from the stems of the grasses and grains

is

made
con-

up

of long slender bast-fibres knotted or thickened at

regular intervals (Fig. 41, a).

stricted at these points

The

central canal

is

and

fine pores are present, passing

through the wall.

tissues
are,

The

characteristic structures of such

cells

however, the

from the epidermis,

flat

and of somewhat variable length, with thick walls and

serrated edges (Fig. 41, c).

In straw pulp a third type of

cell is present,

derived
oval,

from the internal pith

layer.

These are

large,

thin-walled cells with rounded ends (Fig. 41, 6).

Esparto

or alfalfa grass {Stipa tenacissima)

may be

distinguished

from straw by the absence of these pith

smaller size of the bast

cells,

and by the

and epidermal elements.

REFERENCES.
Cross, C. F., and Bevan, E.
J.

Text-book of Paper-making.

London, 1900. Herzberg, W. Papierpriifung.

BerKn, 1902.

WmiNEY, W.
ination of

R.,

and Woodman, A. G. The Microscopic ExamPaper Fibres, Technology Quarterly, XV, 1902, 272.

CHAPTER

IX.

THE MICROSCOPE IN MEDICINE AND SANITATION.

I.

The Microscope

in Biology.

^The microscope bears

that

to biology

much

the

same

relation

the

balance

bears to quantitative chemistry.

It is the

fundamental

instrument without which a true science of living things

would be almost impossible.

The

invention of the
of

compound microscope and

the

construction

simple microscopes of high power at

the middle of the seventeenth century stimulated a dozen

observers to the study of the finer structures of plants

and animals.
in

Leeuwenhoek discovered
invisible

in

water and

decomposing organic matter a teeming world of minute

life

forms of
English

to the

naked
the

eye.

Hooke, the
structure
of

botanist,

made
the

out

cellular

plants in his examination of cork.

Malpighi and Swam-

merdam
It

figured

microscopic details of the insect

success.

body with marvellous patience and

was
only,

however, after the perfection of the

achromatic objective about 1835 that these early labors

could bear abundant
fruit.

In 1838-39 Schleiden and

Schwann developed
their

that great generahzation which bears

all

name, the doctrine that

plants

and animals
104

are

THE MICROSCOPE IN MEDICINE AND SANITATION.

built

105

up

of minute individual masses called cells.

Quickly
cells

the essential identity of the substance

forming the

was postulated.

The microscope

thus grounded anatomy

upon the firm basis

of the cell theory;

and to-day

cytology,

the study of the minuter structure of the cell

itself,

forms

an independent subject of ever-increasing importance.

Next

it

was discovered that the

cell is

not only the unit

living

of adult structure, but thd

form

in

which every

organism originates from

the course of

its

parent.

In the study of
cell
is

development by which the single

transformed into the mature organism, carried out by

Van Beneden, and their compeers, embryology came into being. Upon the anatomical and
KoUiker,

Hertwig,

embryological unity thus demonstrated under the microscope,

the

doctrine of

evolution,

the

most important
century,

scientific

contribution of

the

nineteenth

was

largely founded.
2.

The Microscope

in Medicine

and Sanitary Science.

Of

all

the branches of biology, none owes

more

to the

compound microscope than the study

of disease.

The

conception of the body as a complex of protoplasmic

cells,

-whose normal cooperation was the condition for

good health, led easily to the conclusion that disease

might arise from the deranged functioning of the
dividual
cell.

in-

Cellular pathology,

associated

with

the

name
in

of Virchow, led along this line to great advances

the

knowledge of those diseased conditions which

activity of

arise

from the abnormal

the

living

proto-

plasm.

Another group of maladies due

to the

invasion of

io6
parasitic

ELEMENTS OF APPLIED MICROSCOPY.

plants

and animals, the epidemic plagues, fevers, and pestilences, was comprehended only after their exciting causes were discovered by the microscope.

The
first

bacteria, observed

by Leeuwenhoek
by Ehrenberg

in 1680, were

carefully studied

in the

wonderful

decade of progress which followed the perfection of the

achromatic objective.

Twenty

years later Pasteur estab-

lished the casual relation of these

minute fungi to disease;

and

in the next quarter of a century the organisms caus-

ing anthrax, tuberculosis, typhoid fever, diphtheria, and

cholera were discovered.

These bacterial

parasites are

studied largely by culture methods;

is

but the microscope

causing

also

indispensable.

The animal organism

artificial

malaria cannot be grown on

media, and our

whole knowledge of
the
first

it

depends on optical methods.

In

few years of the twentieth century, the

brilliant

researches which revealed the parasites associated with

smallpox and scarlet fever were carried out with the

microscope alone.

The

researches into pure science which have led, and

are leading, to such advances in pathology

and

parasi-

tology cannot, of course, be treated here in detail.

Cer-

tain applications of the microscope to the study of the

body

fluids

and

of food material have, however,

clinical diagnosis

become
in-

a routine part of
spection service.
cal

and the sanitary

These technical applications of medi-

microscopy come legitimately within the scope of our

work;

and a few

typical

methods

for

the

study of

pathologic conditions and of parasitic invaders will

now

be briefly considered.

THE MICROSCOPE IN MEDICINE AND SANITATION. 107

3.

The Examination

of Urine.

Many organic diseases

etc.,

affect materially the composition of the urinary secretion. Its

amount, reaction, and

specific gravity are significant,

and chemical tests for sugar, albumin,

value.

are of

much

The
is

presence

of

certain

important

organized

elements

determined under the microscope.

The

precipitate
first

which forms in any urine on standing

must be

concentrated by sedimentation, or better,

to a

by the use of the centrifuge, transferred by a pipette

clean slide, covered,

and examined with the high power.

where evaporation
is

The edges
place,

of the preparation,

taking

should be avoided, since the crystals which form

here are not characteristic.

The

principal

objects

which

may

be found in an

examination of urinary sediments are crystals of certain

products of metabolism, red and white blood- cells, epithelial cells,

and

tube-casts.

The

inorganic salts precipi-

tated

may be

diverse in character, including uric acid

(clusters of

rhombic prisms and whetstone forms), acid

urates
ing),

(amorphous, granular masses soluble on warmcalcium oxalate (small octahedral crystals whose

diagonal planes look like the edges on the back of an

envelope),

and ammonium-magnesium phosphate (long

prisms with bevelled edges,

crystals).

known

as the cofhn-shaped

(See Fig. 42.)

In hyperemia, acute nephritis,

and some other conditions, red blood-corpuscles appear

in

the urine

as homogeneous, yellowish discs 7.5

ji

in

diameter, sometimes distorted to a globular

elled

form or

shrivarbi-

and crenated almost beyond recognition.

is

An

trary line

drawn between "normal"

cells

retaining


io8

ELEMENTS OF MPPLIED MICROSCOPY.

and abnormal
colorless

their red color

and crenated

cells.

The

other formed elements of the blood, the white cor-

puscles (polymorphonuclear leucocytes), are larger bodies,

more or

less spherical

and possessing

several nuclei;

the

Fig. 42.
cells.)

urates,

Urinary Sediment in Catarrh oe the Bladder. (Acid ammonium-magnesium phosphate, leucocytes and epithelial
(After

Ultzmann and Hofmann.)

200 diameters.

latter

may

be brought out clearly and differentiated

epithelium by allowing dilute

from the

cells of the renal

acetic acid to

run under the cover-slip when the nuclei of

cell

the two types of

may be made out.

The

characteristics

THE MICROSCOPE IN MEDICINE AND SANITATION. 109

of the white corpuscles will

be treated somewhat more fully

is

in the

next section;

but their presence in urine

of

much significance since it is these cells which mainly make up 'the whitish pus discharged from inflamed
surfaces.

When

the walls of the genito-urinary tract are

cells

seriously

affected,

detached from

its

lining

epi-

thelium will be noted in the urine.

of

The appearance
in Fig.

some

of

these

elements

is

indicated

42.

Finally,

most renal diseases are characterized by the dekidney of an albuminous

posit in the fine tubules of the

substance which coagulates to form minute twisted tubes

(Fig. 43).

These

casts

found in the urine

may

be clear

and hyaline or

may
cells

be rendered granular by the presence

of disintegrated epithelial cells or

may

contain undecom-

posed epithelial

or pus cells or fat globules or bacteria

according to the^ special pathologic condition from which

they arise.
sharply
So-called

waxy

casts are denser

and more an

marked than any other type and

indicate

advanced stage of renal disease.

4.

Examination of Blood.
plasma containing

^The

blood

is

a colorless
types

fluid or

cells of several distinct

whose number and relative proportions vary in

physiologic
of

many
and

and pathologic
to

states.

Few

clinical tests are

more value

the physician both in diagnosis

prognosis than the examination of this

the microscope.

body

fluid

under

For the study of the various types of

cells the

blood
of a
deli-

must be dried and stained;

good blood smear for staining
cacy.

and the preparation

is

a matter of some
the

The blood may be obtained from

lobe of


no
the ear

ELEMENTS OF APPLIED MICROSCOPY.

by pricking with a
washing
sterile

lance or glover's needle

after carefully

the surface' of the skin.

small

drop should be caught on the edge of a clean

slide,

which

Fig. 43.

lar casts, leucocytes,

Urinary Sediment in Acute Bright's Disease. (Granuand epithelial cells.) (After Ultzmann and
200 diameters.

Hofmann.)

is

then brought in contact with the surface of a second

clean slide.
at
its

The upper glass is drawn along the lower one an acute angle so that the blood-drop escapes beneath
edges and
is

spread over the lower slide in a thin

THE MICROSCOPE IN MEDICINE y4ND SANITATION,

even smear.
staining.

After drying in the air this

is

ready for

One
is

of the

most satisfactory

differential blood- stains

Wright's modification of Leishman's method, described

the Journal of Medical Research for January,
p. 138).

in

1902

(volume VII,

The somewhat

complicated pro-

cedure for making this stain and some discussion of

the philosophy

of its action must be sought

student will find
is

in the original

paper; but

the

its

application easy.

The
by

dried film

covered with the solution of the dye in

is

methyl alcohol for one minute and water

drop,
until

added, drop

the

mixture becomes

semi-translucent

and a yellowish metallic scum forms on the surface.

This mixture
for

is

allowed to stand on the preparation

off in distilled
its

two or three minutes and then washed

till

water

the film has a yellowish or pinkish tint in

thinner portions.

The

slides thus stained, dried,

and mounted

in

balsam

should show the various types of blood- cells beautifully

differentiated.

The

red corpuscles, or erythrocytes, are

orange

or pink in color
cells

and about

7.5

fi

in diameter.

The white
to

may

be divided into three classes accordtheir nuclei

stain.

ing to the relation the

constituents

which

and granules bear

eosinophiles or

of the

The

oxyphiles are cells containing matter which takes such

acid stains as eosin.

Stained with the Wright stain, these

lilac- colored

appear faintly blue with a dark

nucleus

and numerous large reddish granules.

the size

They

are double

of the erythrocytes.

The

basophiles are cells

varieties.

which take basic stains and are of two

Small

'

112

ELEMENTS OF APPLIED MICROSCOPY.

or

basophiles

lymphocytes are about the

size

of

the

red corpuscles, robin's egg blue in color with a round

nucleus and a few fine dark-blue granules. mononuclear basophiles are three times the size Large

dark-blue

of the red cells

and pale blue with a large oval blue nucleus.

eosinophiles
are

The
the

third type, the neutrophils, are intermediate between

red-granuled

and

the

blue-granuled
twice
the

basophiles.
size

They

polymorphonuclear,
blue, with

of

the erythrocytes, and

one or more

darker lilac-colored nuclei of a twisted shape and numerous

medium- sized
bodies

granules

of

reddish-lilac

color.

Blood-plates are almost always seen, purplish rounded

or oval
one-third
the
size

of

the erythrocytes,

with irregular edges and fine blue mottlings.

logic conditions

In patho-

numerous other

cell

elements

may

be

present.

Most

of these cell elements are seen in Fig. 44.

is

Such a stained preparation as that described

the study of the structure of the blood-cells

used for
for the

and

determination of the relative

various types.
is

number

of white cells of the

it is

differential

blood count, as

called,

made by counting

several

hundred white

cells

and
de-

calculating the percentage of each form.

For thus

termining the absolute number of the blood corpuscles the

apparatus called a haemocytometer

sists of

is

designed.

It con-

a glass slide containing a cell of

of o.i

known

diameter
is

and a depth

mm., on

the bottom of which

off

ruled

a micrometer scale marking

side.

squares 0.025

"^^i-

on a

The
the
cc.

blood

is

diluted, in a special pipette constructed

for

purpose,

with

100 parts of Gower's solution

g.

(100

water, 15 cc. acetic acid, 6

sodium sulphate),


THE MICROSCOPE IN MEDICINE AND SANITATION. 113

when red
cells

cells are
is

to

be counted.

For counting white

the blood

diluted with 10 parts of

^%

acetic acid
cells

tinted with gentian violet,

which destroys the red

and

faintly stains the white corpuscles.

Wide

variations of the blood elements, both in absolute

in proportions, occur in

numbers and

many normal and

Fig. 44.

Blood-cei-ls (Wright's Stains). (Photomicrograph by W. C. Greene, Harvard Medical School.) (Erythrocytes, large basophiles, polymorphonuclear neutrophiles, and blood plates.) 700 diameters.

abnormal conditions.

The blood of the

adult

human being

should contain 5,000,000 erythrocytes per cubic millimeter

in the

male and 4,500,000 in the female and 3000 to

of

10,000 white blood-cells,

which 70-72% should be

polymorphonuclear neutrophiles, 25-30% basophiles, and

1%

eosinophiles.

In anaemia the red

cells

may

fall to

360,000 per cubic millimeter and in other conditions they

may much

exceed the normal.

Leucocytosis, or the in-

114

ELEMENTS OF APPLIED MICSOSCOPY.

accompanies
all

crease of white cells,

acute inflamma-

tions as well as certain other conditions

and may

affect

any one type of

it

cells

or

all

of them.

In inflammations

is

the

polymorphonuclear

neutrophiles

which

are

increased; in trichinosis, on the other hand, the eosinophiles

5.

may

reach

50%

of the total.

The Detection
are

of the Malarial Parasite.

Blood

ex-

aminations
malaria.
to the

also

undertaken for the

diagnosis

of

The Protozoon

malarial parasites gain entrance

body from the

bite of

an infected mosquito

in the

form of minute spore-like bodies which penetrate the

red corpuscles and there assume a crescentic form.
the organism

Later

becomes amoeboid, sends out blunt pseudo-

podia,

and develops nuclear granules.

As

it

matures,

it

occupies the greater part of the red corpuscle and finally

begins to divide, appearing in a rosette form, at last separating

and discharging

into the blood stream granules

like those

with which the cycle began.

Blood

may

be examined in

its

fresh condition for the

malarial parasite, but better preparations are

obtained
yields

by

staining.

The Wright-Leishman method

the

ad-

mirable

results,

cytoplasm

of

the

parasite

being

stained blue
lilac

and the nuclear granules a color varying from

through red to almost black.

of
blood-cells

The

general appear-

ance

containing the parasite of tertian

is

malaria,

HcBmamceba vivax,
;

indicated in the two upper

fields of Fig. 45

the lower field shows a later flagellated

stage in the flood stream.

in the examination of

Only considerable experience normal blood will enable the ob-

server to recognize the invading organism with certainty.


THE MICROSCOPE IN MEDICINE MND SANITATION. 115

The presence

in the

blood of free "pigment," probably

stainable material
sites, is

from decomposed blood-cells or para-

characteristic of malaria.

Fig. 45.

The Malakiai Parasite

Spitta.)

(Stained). 1000 diameters.

(After

Slater

and

6.

The Examination of Spufum for Tubercle

Bacilli,

and the Cultural Diagnosis of Diphtheria.

Besides

the

II

ELEMENTS OF

/IPPLIED

MICROSCOPY.

examination of blood smears for malaria the bacteriologist of

a Board of Health

is

expected to

make

diagnoses

of diphtheria

and

tuberculosis,

which are of special im-

portance on account of the necessity for prompt treat-

ment

of the former disease

and on account

of the difficulty
latter.

sometimes experienced in recognizing the

Both

maladies are due to the presence of bacteria, very minute

rod-shaped fungi which

first

gain lodgment in the respira-

tory or alimentary passages.

In the case of consumption or pulmonary tuberculosis

the bacteria are discharged in

enormous numbers

in the

sputum and may

there be detected without great difficulty.

When
teria,

stained with an anilin-dye the tubercle bacilli are

not decolorized by dilute sulphuric acid as are most bac-

and

this

property

is

made

the basis for a simple

method
is is

of differential staining.
glass slide

thick smear of

sputum
This

made on a

and dried over the flame.

it

stained for two minutes, heating until

steams, with

Ziehl-Neelsen's carbol fuchsin (i

gram

basic fuchsin in

ID

cc.

95%

alcohol mixed with 90 cc. of a

5%

aqueous

solution of phenol).

The

slide is

then washed under the

tap and immersed for two minutes in a solution of 3 parts

of hydrochloric acid in 100 parts of

95%

alcohol.

It

is

then washed again and counterstained for half a minute

in a dried,

1%

aqueous solution of methylene blue, washed,

and mounted.

The

slender tubercle bacilli should

be stained bright red with fuchsin, while other bacteria

and the leucocytes and other

colored faintly blue.

cell

elements present are

In

all

work with

bacteria, the -fV-inch objective

must

THE MICROSCOPE IN MEDICINE /IND SANIT/ITION.

n?

be used with a drop of cedar- oil between the lens and

the cover-glass, or the slide,
if

no cover-glass be used.

The Abb^ condenser

terial

is

also a necessity for lighting bac-

preparations.

The

diagnosis of diphtheria

is

somewhat more comand involves cultural

plicated than that of tuberculosis

as veil as microscopical
at the
is

methods.

The organism

occurs

back of the throat and material for examination

obtained by rubbing a sterile swab of cotton over the

whitish patches

which appear on the pharynx and fauces.

This swab

is

then passed over a slanting surface of coag-

ulated blood-serum
to

upon which

after incubation for six

twehe hours
of the
is

at the
bacilli

body temperature minute pearly

will

colonies

appear.
sterile

portion of the

growth

then removed with a

loop of platinum

wire and spread over a clean cover-slip in a drop of

water.

The

cover thus prepared

is

dried, fixed

by

pass-

ing three times through the flame,

and stained
(30

for ten of

minutes

with

LoefHer's

methylene blue

cc.

a
cc.

saturated alcoholic solution of the dye

of

mixed with 100

a Vio.ooo aqueous solution of caustic potash).

drying,

After

washing,

and mounting

in

balsam, the speci-

men, in cases of diphtheria, will show the very variable

rods characteristic
of

the diphtheria bacillus,

many

of

them cigar-shaped or club-shaped, and often exhibiting

the peculiar cross-barred appearance
46.

indicated in Fig.

Sometimes

these

organisms

will

be

seen

mixed

with spherical bacteria arranged

the
streptococci.

in

pairs

and chains,

In negative

cultures

only cocci are

foimd,

as a rule, although

other bacilli,

more or

less


ii8
closely

ELEMENTS OF APPLIED MICROSCOPY.

resembling
the

diphtheria

organism,

may

be

present.
7.

The Serum Reaction

most
important
progress
in

in Typhoid Fever.
results

One

of

the

practical

of

the

rscent

marvellous
of

the

study of

the

phenomena

immunity has been a method

of

for the clinical diagIt

nosis

typhoid fever and certain other diseases..

Fig. 46.

Diphtheria

Bacilli
brook.)

(Methylene Blue).
Diagrammatic.

(After

Wes-

has been shown that

when

the animal

body

is

invaded

by a
cases

parasitic micro-organism, the cells

produce in some
of

antitoxins

which neutralize the poisons

the

microbe, and in other cases anti-bodies of another type

which have a
itself.

specific destructive action

on the

parasite

Sometimes the defensive secretions dissolve the

THE MICROSCOPE IN MEDICINE AND SANITATION.

foreign cells;

"9
to-

sometimes they cause them

to

clump

gether in masses

and

to settle out of the fluid in

which
in-

they are suspended.

In the blood of an individual

fected with typhoid fever, bodies of the last type,

as agglutinins, are present;
specific for that particular

known
is

and

their

clumping reaction

micro-organism.

It is true that

the blood of a

normal individual
bacilli,

may

contain substances

which agglutinate typhoid

patient has the

but that of a typhoid

it

power

to

such a high degree that

will

produce the same effect

dilution.
case, a

much more
is

rapidly and in high

In making the diagnosis of a suspected typhoid

taken from the lobe of the ear
to clot in a small test-

sample of blood

in the usual

manner and allowed

tube so that the corpuscles are separated from the clear

straw-colored serum.

drop of serum

is

then mixed

on a

slide

with 40 drops of a fresh broth culture of the

typhoid bacillus.

The mixture

is

covered and examined

under the
bacilli

^^g^-inch

oil-immersion objective.

At

first

the

may be

seen, after careful focussing, as

minute trans-

parent rods, singly, or in pairs

and short

chains,

moving

rapidly about across the field of the microscope.

tinins

If agglu-

be present, the bacteria become motionless and clump

together in masses, sometimes large

to the

enough

to

be

visible

naked

eye.

If the reaction is not obtained

with

a jV dilution, another drop of serum

if

may

be added, and
should

this fails, again, another.

The

latter dilution, xVi

if

clump the

bacilli in fifteen

minutes

the case examined

be typhoid fever; with a ^V dilution an hour's time may be needed. The test may be made quite as well with dried
blood after diluting with the proper

amount

of water.

I20
8.

ELEMENTS OF /IPPLIED MICROSCOPY.

Th3 Microscopic Examination
of
is>

Pork

for Trichina.

The disease
Trichina
is

known

as trichinosis

like diphtheria

and

typhoid fever caused

by a

parasitic

micro-organism,

although the parasite belongs to a very different group.

a minute worm, barely visible as
a. speck

to the

naked

eye,

which bores
itself in

its

way

into the muscles of swine

and there encysts

a calcareous nodule. If such pork

is

be eaten, imperfectly cooked, the limy cyst

the

dissolved;
of their

worms emerge and reproduce, and a myriad

progeny penetrate the tissues of the body, causing high

fever and, sometimes death.
this crisis, the

If the patient recovers

from

worms

encyst themselves in the muscles

serious difficulty.
is

and produce no further

In America, trichinosis

rare in
it,'

man, although some

is

2%

of swine are affected with

because pork

more

or less thoroughly cooked before eating.

In Europe,

however, where pork products are eaten almost raw, the

danger

is

serious

and must be met by preventing

the

distribution of trichinous swine flesh.

Elaborate governinstituted in

ment systems

of.

meat inspection have been

in

many
States

countries;

Prussia, for example,

this purpose.

over 25,000

officials are

employed for

In the United

the

Department of Agriculture for some years

maintained a bureau of meat inspection which examined

all

pork intended for foreign export and

interstate

com-

merce, employing a large corps of microscopists.

Samples of pork

to

be examined for Trichina are taken

from the diaphragm or other muscles and cut in pieces about an inch by half an inch in size. One such piece
is

then placed between two slides of heavy glass, mounted

THE MICROSCOPE IN MEDICINE AND SANITATION. 121

in a

frame and provided with screws, by which they can

be pressed together, thus reducing the meat to a thin

transparent layer.

The frame
fit

bears on

its

under side

two ridges which

into grooves in a special corrugated

for this

stage of large size

on the microscopes designed

purpose.

Under a very low

objective, of i|-inch equivais

lent focus, the

slid

compressed frame
its

then examined, being

along until

whole width has been covered, and then

a

shifted to the next groove, so that

new

examined.
in five

In this way the whole sample

field may be may be viewed

minutes by a trained observer.

is

Under the microscope the pork

seen to be

made up

of the long cylindrical cross-striated fibres of voluntary

muscle, and
here

if

Trichina be present the cysts will be seen

and there as spindle-shaped, whitish bodies.

In

sections of trichinous

pork prepared after paraffin im-

bedding with the microtome, the

coiled
9.

worms may be seen

in Fig. 47.

up

inside the cysts, as

shown

The Microscopy of Drinking Water.

^The
its

sanitary

quahty of water depends primarily upon the presence

or

absence of disease

germs,

generally introduced

in

sewage.
is

The most important

evidence as to

character

therefore obtained

by the bacteriological and chemical

analyses,

which indicate with great dehcacy the presence

sewage pollution.
often

of minute traces of

microscopical
of

examination may,
value
cells,

however,

add information

by showing the presence of starch-grains, yeastfragments of vegetable tissue, and certain Infusoria

characteristic of
iar to

decomposing material; organisms pecul-

a particular pond or stream have som^tirnes proved


122
of

ELEMENTS OF APPLIED MICROSCOPY.

its

importance in the detection of admixture of

water

with that from some other source.

The

great importance
lies,

of the- microscopical examination of drinking-water

however, in

its

appUcation to the study of those organisms

tastes

which produce
tastes

and odors

in

reservoirs.

Such

and odors are

in almost all cases

due

to the mul-

FiG. 47.

Trichina.

(After Hager-Mez.)

50 diameters.

tiphcation of microscopic plants

to the Algas

and animals belonging

control
of

and the Protozoa; and the study and

of these organisms

depends upon the systematic use

the microscope.

considerable concentration of the sample must, of

course, precede the microscopical examination of drink-

ing-water;

and

this is usually

accomplished by

filtration

through a layer of

fine

sand.

The
of

Sedgwick-Rafter

method, in routine use by the State Board of Health and

the

Metropolitan Water Board

Massachusetts,

the

THE MICKU^LUft IN MtUIUNb /INU

Water Department
of
:

i/INI 1 .1 1 lUN.

'^>J

Brooklyn, and

other

sanitary

authorities, is as follows

measured quantity of water,

through perhaps a quarter
circle of bolting-cloth suptall
is

half a liter or a liter,

is

filtered

of

an inch of

fine

sand on a

ported by a perforated rubber stopper in a

funnel.

The sand with

into a test-tube,

the organisms collected

on

it

dropped

shaken up with a small volume of water,

settle for

and allowed to
poured
to
off

a moment.

The water

is

then

and a second portion of wash-water serves

all

remove almost

the organisms from the sand.

The
some
in say

two washings mixed together are then

convenient voluine, lo or 15 cc,
10
cc.

made up
liter.
is

to

and thus one has

the organisms originally present in a

One

cubic centimeter of the concentrate

then placed

in a cell, i

mm. deep and

1000
is

sq.

mm.

in area.

A special

type of ocular micrometer

used with a square ruled

upon

it,

and the lenses and draw- tube are so adjusted

that the square shall coincide with i sq.

mm. on
counted,

the stage.

Ten
then

or twenty fields in different parts of the cell are

examined and the

organisms

when a

simple calculation enables one to

present in
i cc.

compute the number

of the original sample. of micro-organ-

By

this

method the kind and number

isms present

may

be easily determined, the cause of

existing trouble detected,

culties predicted.

and the prospect of future

efficacy of

diffi-

The

methods

of purificain

tion

may be

tested

and the source of trouble located

where
it

the particular part of a supply

exists.

few of the most serious odor-producing organisms

are figured in Fig. 48.

At 4

is

AnabcBna, the Blue-green


124

ELEMENTS OF APPLIED MICROSCOPY.

city of Springfield,

Alga which has cost the

Mass.,

many
a

thousands of dollars by the production in Ludlow Reservoir of the

"pig-pen" odor.

At

is

Asterionella,

Diatom which caused

the geranium odor in the water of

Fig. 48.

Micro-organisms of Drinking-water.
Whipple.)
I, i, 3.

(Redrawn from

4, 5.

300 diameters. Asterionella. 6. Synura. Anabaena. 7. Uroglena.

Brooklyn, N. Y.

The

Protozoon, Synura, at 6 was the

author of the cucumber taste in the Boston supply,

and
of

its ally,

Uroglena, at

7, is

perhaps the worst offender

all,

having caused trouble in some 44 supplies in Massa-

chusetts alone.

THE MICROSCOPE IN MEDICINE /iND SANITATION, ti^

REFERENCES.
Cabot, R. C.
1903.
Clinical

Examination of the Blood.

of Disease.

New New

York,

Cabot, R. C.
1899.

The Serum Diagnosis

Protozoa and Disease.

York,

Clark, C. EwiNG, J.

J.

New

York, 1903.
Phila.,

Clinical Pathology of the Blood.

J.

1903.

McFarland,
MuiE,
R.,

Text-book upon the Pathogenic Bacteria.

Phila., 1900.

and Ritchie, J. Manual of Bacteriology. Amer. by Harris, N. M. New York, 1903. Oertel, T. E. Medical Microscopy. Phila., 1902. PuRDY, C. W. Practical Uranalysis and Urinary Diagnosis.
ed-,

Phila., 1895.

Tyson,

J.

Guide

to the Practical

Examination of Urine.

Phila., 1902.

Whipple, G. C.
York, 1899. Wright, J. H.

The Microscopy

of Drinking-water.

New

of Blood Films

Rapid Method for the Differential Staining and Malarial Parasites. Journal of Medical

Research, VII, 1902, 138.

CHAPTER

X.

FORENSIC MICROSCOPY.
1.

The Microscope in Law.

li'"e

In a court of law where

in
to

questions of

and death may hang upon the nature

study the minuter
identification
of

of a blood- stain or the genuineness of a document, the

microscopist
structures
of

is

often

called

disputed

objects.

The

persons by the anthropometric system of Bertillon

may

be supplemented to advantage by a detailed examination

of the ridges

upon

the fingers.

The

detection of

human
be

blood

is

greatly

facilitated

by microscopical methods.

In sexual cases the presence of spermatozoa

easily

may

determined under the microscope.

Low

powers

aid in

the study of bullet-wounds with respect to the

distance

and

direction
etc.

from which the shot was

fired,

the
as

powder used,
regards
the

The examination
writing

of documents,

paper and

utensil

used,

the

presence of erasures or alterations,

istics

and the

characterfield

of

the

handwriting,

is

an important

for

microscopy.
2.

The Examination
is

of Blood Stains.

^Frequently the
minute
126

expert
stains

asked to decide as to the character of bloodclothing, furniture, etc., stains often of

upon


FORENSIC MICROSCOPY.
size

127

and sometimes

of considerable age.

Two

problems

are

involvedfirst, as

to the presence or absence of blood

its

in the

specimen presented, and second, as to

nature,

whether
general

human

or animal.

The

presence of blood in

may be

settled pretty easily

by four methods
one
microspectro-

one

chemical,

one

microchemical,

scopical,

and one purely microscopical.

The chemical

test consists in the

application to the stain of an alcoholic

solution

of

gum

guaiacum, which yields a blue color

reaction

on the further rddition of aerated turpentine.

test

The microchemical
globin, the

depends on the fact that haemo-

complex albuminoid substance characteristic

of the

red blood-corpuscles, yields on treatment with

acetic acid

a crystalline iron

salt,

hasmatin,

whose

triple

chloride

is

known

as hsemin.
is

In making

this test the

blood,

if

in

a dry condition,

placed on a slide and geritly

warmed
rate

in glacial acetic acid.

This

is

allowed to evapo-

and replaced by a 0.07% solution of sodium chloride,

its

which in

turn

is

carefully evaporated without heating

the slide above 60 C.

Another drop of acetic acid

is

added and the

boils.

slide is this

time heated so that the liquid

is

After evaporation
if

it

examined under the

^-in.

objective;

blood be present, crystals of haemin (or

flat

haematin trichloride) will appear as minute

rhom-

boids often superposed in the form of a cross.

They

vary from yellow to reddish

0.00SHD.02

brown
test is

in color

and from

mm.

in size.

This

a delicate one, serv-

mg

to detect

from 0.05-0.15 mg. of dried blood, and even

very old material gives satisfactory results, as in the classic

case of the blood of the

murdered Kotzebue, detected

128

ELEMENTS OF /tPPLIED MICROSCOPY.

on the papers which lay upon

sixty years after his death

his desk.

For

examination

by

the

microspectroscope,

fresh

blood-stains are treated with distilled water and older

specimens with weak acetic or

in a small glass cell

citric acid.

When

placed
to be

and examined by the methods

described in the next section, the characteristic spectrum

of haemoglobin or one of
its

derivatives will be apparent.

The
and

direct microscopical examination of blood-stains at

aims simply
serves

the

detection

of

the

red

corpuscles
that

to distinguish

mammalian blood from

of birds, the

three

tests

above outlined being general

Dried

for the haemoglobin of all red-blooded animals.

and and

clotted

blood

must

first

be treated with

some

solvent which shall attack

set

the

albuminous coagulum

free

the

blood-cells.

Numerous media have

solution of potassium

been suggested, of which a

hydrate
is

33%

one of the simplest, and Ranvier's solution

2

(potassium iodide,
of iodin, 100 parts)

parts;
is

saturated aqueous solution

one of the best.

In any case a

bit of clot or the scraping

from a

stain is covered with

the solvent
scope.

and placed

in a hollow slide

under the micro-

Sometimes

after

a few minutes, sometimes only

after days,

the opaque mass becomes clearer and the

individual cells appear as circular, non-nucleated discs

7.5
//

in diameter.

Under most conditions the

discs

appear somewhat biconcave, with a thickened ring about

the edge.

In the

original process of drying or in the treatment

with a solvent, the blood-cells

may

be so distorted that

FORENSIC MICROSCOPY.
their

129

recognition

is

not easy.

In water, for example,

the corpuscles swell

up

to a spherical form.

The comlitera-

monest objects which

may be mistaken
instances
of

for blood-cells

are the spores of certain plants,

ture

and medico-legal

contains

ludicrous

erroneous expert

opinions based on such findings.

tion carefully

With a good preparaof fresh blood,

compared with specimens

such errors can easily be avoided;

puscles of birds are of oval shape
nucleus, blood- stains of

and, since the cor-

and show a

distinct

avian origin

may

be excluded.

The blood
that of

of other

mammals can be

differentiated

from

man

only by the size of the

cells,

and the

differ-

ences are so slight that a certain result can seldom be

reached.

The blood

of the goat has cells less

than 4.5/1

in diameter, while the corpuscles of the horse

and cow

and pig are under 6

rabbit, closely

mouse,

cat,

n; but the corpuscles of the dog, and other domestic animals very

resemble those of

human

blood.

Such

differ-

ences of one or two micromillimeters cannot be relied

upon, since even in the fresh blood of a living animal

some variation occurs and distorted dried specimens

are

much more

dubious.

Dr. E. L. Walker has pointed

out that the white blood-cells of different

mammals

differ

much more widely than do

study of these bodies
microscopist.

the red corpuscles;

and the

may

furnish valuable aid to the

The

crucial test for blood-stains,

and the only method

by which the blood of

man may be

definitely distinguished
specific

from that of other mammals, depends upon a

biologic reaction

similar to that

which

is

used in the

11

13

ELEMENTS OF APPLIED MICROSCOPY.

test for

serum

typhoid fever.

If

human blood-serum
which

be

injected into the

lymph

spaces of a rabbit, there appears

precipitates
this reac-

in the blood of the rabbit a substance

certain albuminoids in the blood of tion serves to distinguish

man; and

human

blood from that of the

domestic animals with certainty.

puscles of apes

Only with the

of the
test,

cor-

and monkeys does the serum

immuwhich

nized rabbit react in a similar manner.

ordinarily bears the
it, is

This
first

name

of Bordet,

who

suggested

not applied under the microscope, but in small tubes

is

in

which the precipitation

3.

detected macroscopically.
Its

The Microspectroscope and

and
in

Use.

In

testing

blood-stains,

many

other fields of microbiology

aid.

and microchemistry, the spectroscope may furnish

As
as

ordinarily used,

the instrument consists of a tube

of a microscope

fitting

on the eyepiece

and

containing,

shown

in Fig. 49, a triangular prism of flint glass

light

which disperses white

into

its

constituent colors

and two prisms

tion,

of

crown

glass set in the opposite direc-

which serve
first

to counteract the refraction produced

by the

prism.

The

principle

is

the

same

as that

of the achromatic objective, but whereas in

the latter

the angles are arranged to produce a net refraction with

neutralized
dispersion,

the

prisms of the spectroscope

yield a net dispersion with practically

no

refraction.

The

special

type

of eyepiece

used with the micro-

spectroscope contains a diaphragm which only allows a

longitudinal
slit is slit

of light to pass,

and the width

of the
for use

regulated by a set screw, being cut

down

to half a millimeter or less.

When

the detachable prism


FORENSIC MICROSCOPY.
'

131

tube

is

placed in position and the

field well lighted,

its

spectrum should be seen lying with

colored bands at
If the

right angles to the slit in the ocular.

lines are oblique, the prism tube
it

spectrum
until

must be rotated

bears the proper relation to the sHt.

Reference to Fig. 49 will show also a lateral tube connected with the spectroscope, into which rays of light

Fig. 49,

The Miceospectroscope.

(Bausch and Lomb.)

from outside are projected by a mirror, and a prism

by which these rays are

reflected

upward.

This

is

used

when
light

it

is

desirable to

compare the spectrum of sunof the

with that of some object upon the stage

microscope.

When

properly adjusted the two

light passing

spectra,

produced respectively by

up through the

microscope and by that reflected through the side tube,

will

appear to

lie

side

by

side.

132

ELEMENTS OF APPLIED MICRCSCUFY.

sensation of light
is

The
tions, '

produced by the ether vibra271

whose length lies between

and

155
10,000,000

io,ooo,cxx>

of an inch.
it

When any

solid

body

is

gradually heated

becomes luminous,

first

emitting the longer red rays

later

at a rate of

458 million of millions per second and

the rays of other colors as well, culminating with the violet

rays,

whose

rate

is

727 million of millions.

At

this

point the
all

body has reached a white

off.

heat, since rays of

colors are given

If

now

the white light from such

an incandescent body be passed through the spectroscope,

it

is

broken up, through the different

refrangibility of

the rays of different amplitude, into a continuous spectrum

in

which

all

the rainbow colors appear merging into each

ether.

An

incandescent gas, on the other hand, profield

duces a line spectrum, most of the

being dark, with

here and there narrow bright lines whose

number and

position are characteristic of the particular substance.

The

color of objects

is

due

to their property of transIf

mitting or reflecting rays of a certain amplitude.

white light be passed through certain

gases

solids, liquids, or

below

their

point

of

incandescence,

and then

through the spectroscope, the presence of black bands

crossing the spectrum shows that light of certain definite

wave-lengths has been removed.

gas cuts out the

same light-waves which

temperature.

it

itself

produces at a higher

The spectrum produced by

one, like that

sunlight

is

a continuous
solids,

produced by other incandescent

crossed here and there

by dark bands, the Frauenhofer

FORENS/C MICROSCOPY.
lines,

I33

which are due

to the presence of the cooling vapors

of metallic elements in the of

surrounding atmosphere, each

its

which absorbs

the

hght corresponding to

own
con-

bright line spectrum.

sidered the

The Frauenhofer lines may be

shadows of the spectra of the metals

in the

outer atmosphere of the sun.

Obviously

artificial light

should be used for the micro-

spectroscope, since

the Frauenhofer lines

would
solid

intro-

duce

confusing

element.

When

various

and

liquid substances are placed

on the stage

of the micro-

scope, absorption spectra of the

same general type

as

that of sunUght are produced, although the

dark bands

are wider

and

less

sharply defined, this being moire and

of the

more the case as the thickness

is

body

is

increased.

It

therefore desirable to examine layers of varying depth,

this

and

may be done by means

piece of
shall

of a cell

made by cementoff obliquely

ing to a slide a
so that
it

heavy glass tubing cut

be

mm.

deep on one side and 0.5

mm.

deep on the other.

The
ination

application of the microspectroscope to the examof

various

organic

bodies
its

has

not

yet
It

been
has

developed to anything like

possible limits.

yielded interesting results in the study of certain animal

and vegetable coloring-matters;

so far

but

its

chief

use has

been in the detection of blood-stains, since the

of

red

coloring-matter

blood

produces

characteristic

spectra.

Suspected blood-stains which are to be thus

examined

may

be

treated with distilled water until a

is

reddish-brown solution
that

produced, when

it

is

apparent

haemoglobin has gone into solution.

The hquid


1.34

ELEMENTS OF APPLIED MICROSCOPY.

thus obtained,

when placed
with

in a cell, covered

and

exof

amined with the low

oxyhaemoglobin
green,

objective,

shows the spectrum

in

dark bands

the

yellow

and

shown

in Fig. 50 at

4.

If a

drop of

ammonium

sulphide be added, the two bands will fuse into one, giving

70

65

60

5,5

50

45

Fig. 50.

Spectra of Oxyhemoglobin and reduced Haemoglobin.

(After Howell.)

the

spectrum

of

reduced

haemoglobin,

shown
acid

at

B.

The

addition of a

little citric

acid to a solution of oxyhaematin,

haemoglobin produces the spectrum of

with a broad band in the red, a narrow one in the green,

and a very

faint

band

in the blue.

In old blood-stains

the oxyhaemoglobin has been changed to haematin, and

since
this

substance

is

insoluble

in water,
is

no

color-

ation of the liquid appears

when

the stain

treated as
tried;

above.

In such a case a

Httle acetic acid

must be
at

this dissolves hasmatin,

and the solution

once pro-

duces the acid haematin spectrum.

On

adding

am-

monia, the broad band in the red disappears.

Stauas in

Blood-

which the haemoglobin

is

only partially changed

FORENSIC MICROSCOPY.
to

I35

hEematin

(methaemoglobin)

with the combined bands of both.

test is, -of course,

may show The

spectrum

spectroscopic

given by the blood of any red-blooded

animal

It is

extremely delicate and has yielded posi-

tive results
4.

with stains over a hundred years old.

of Finger-prints.

The Study

The
cases,

identification of

individuals, so important not only in criminology but in

a multitude of
facilitated
tillon.

civil

and criminal

has been greatly

of

by the anthropometric system

rests

M.

Ber-

This

upon

the principle that variations in

the proportions of the body,

when

number

of separate

features are measured, are so characteristic that a definite

formula

may

be obtained from them, which shall

differentiate

each individual from any other.

of the

Commonly
of the

the length
left

and breadth

head and the length

middle

finger, the left forearm,

and the

left

foot

form

a primary basis for classification to which other general

characteristics

and

special individual peculiarities

may

be added ad infinitum.

The
to

use of the finer details of the structure of the skin

first

supplement the Bertillon system was

biologist,

suggested

by the distinguished English

prints as a

Francis Galton,

though Sir William Herschel had previously used finger-

means

of personal identification.

Recently

Professor H. H. Wilder's investigations have

great importance
heredity.
If

shown

the

of

these structures in

the

study of

the bulb below

the tip of the finger be

thin
card,

pressed

first

upon a metal surface covered with a

and then upon a paper or

film of printer's ink,

a permanent record will be obtained of the arrangement

136

ELEMENTS OF APPLIED MICROSCOPY.

and furrows
characteristic of that digit, a

of the ridges

record necessarily free from the error which

may

attend

ordinary

anthropometric

measurements.

Galtpn

has

shown

that the pattern of the skin persists

life

unchanged

through
burns,, or

and

is

unaltered in

its

essentials

by

cuts,

any ordinary accidents.

The

variations are so

great that he calculated the chances of identity between

two

single

prints to

be only one in sixty-four

is

billion.

When

ten digits are compared, identification

records, in order to

absolute.
service,

Such

be of any practical
classified;
this

must be readily arranged and

Galton

2
(Redrawn

'
after Galton.)

Fig. 51.

FiNGEE-PRiNT Patteens.

has done

so successfully that, out of a " finger-print direc-

tory" of 2632 persons, any pattern could be located in

three minutes.

In the

first

place, the patterns of the skin

may

be grouped under three heads.

In

all.

cases the

papillary ridges run across the fingers in the vicinity

of the third joint, of the nail in a

and

at the tip they follow the curve

rounded arch.

Sometimes the ridges

between follow the outer ones in a

arch of lessening convexity; this
is

more or

less

even

the arch type

(i.

Fig. 51).

Sometimes the intermediate ridges form a loop running from one side inward to the center of the bulb and then
doubling back again.

Obviously at the opposite side

FORENSIC MICROSCOPY.
of the finger the outer ridges, the loop,

137

and the basal

ridges will cut off

between them a
51);
this
is

triangle,

known
to

as

the delta
the ridges

(2,

Fig.

the loop type.

Finally,

on the bulb may be so twisted as

two
deltas,

form a

complete

circle cutting off

one on either side;

this is the

whorl type
of

(3, Fig. 51).

classification

the

ten digits according to their

for a

arches, loops,

and whorls serves

primary division

of finger-print cards into a

number

of general classes.

Since, however, certain combinations of these digital for-

mulae are
division

must be made; and

much more common than others, further subthis may be accomplished by

by counting the
ridges in the. loops of those
It
is

studying the minuter structures of the finger-print, and

particularly
digits

which show that

structure.

here that the

microscope comes into play, since such details cannot

well be

made

out with the naked eye.

is,

magnification

of only ten or twenty diameters

sirable;

however, generally de-

hence the instrument used must be either a simple

microscope, preferably

mounted on a
of

stand, or a comfield.

pound microscope
5.

of very low

power and very wide

The Examination

Documents.

In
its

the study of

disputed documents the microscope

ble service to the legal expert.

may be of consideraThe material of which

and
water-marks

paper
should

is

made,

its

texture

and

sizing,

first

be noted, the character of the writing instruit

ment and the grooves

cuts

upon the paper,

Inks

if

a steel

pen, being possibly significant.

various chemical methods;

but

may be tested by much may be learned

under the microscope from an observation of the color

138

ELEMENTS OF APPLIED MICROSCOPY.

luster of the deposit

and metallic
taining

formed by inks con-

gums and

of the solid particles of coloring-matter

always present.

Even

the changes of consistency in the

this

same

bottle

on standing may be detected by

method.

Direct evidence of forgery in the shape of alterations,

erasures, or interlineations
is

often furnished

by exam-

ination with a

hand

lens or

low-power compound
sizing

microscope.

Erasures remove the

and loading

material of the paper and leave loose ends of teased-out

fibres

in

which the ink of

later writings

runs

freely.

Marks
in

of preliminary tracings are sometimes apparent of

it

cases

elaborate

forgery.

"When two

lines

cross

each other

may

be of importance to determine which

was made
service,

first;

and

here,

too,

the microscope

is

of

since the

upper Une often shows a widening,

due

to capillarity,

on entering the lower hne, and a narit.

rowing on leaving

The

continuity of the pen fur-

rows of the upper hne

may

also be apparent.

ObUquely
lens, along

viewing the point of intersection with a

the two Unes successively, helps to

hand
it

make
itself,

clear which

one was superposed.

Proceeding
spacing of the
to
lines,

the

handwriting

the
first

general

words, and letters should

the shading,

be noted.

The pen
of

pressure,

the general symmetry

lines are

words and letters, and the firmness of individual

all significant.

caution, in
feebleness,

Temperament manifests energy or reserve. Lack of

itself in

haste or

facUity, physical

and the labored attempt

to imitate, alike pro-

duce tremors which

each other.

may

easily

be distinguished from

FORENSIC MICROSCOPY
All these obvious peculiarities
skilful

139

may

be imitated by a

hand, and

it

is

only more minute characteristics

which serve for the certain identification of handwriting.

2,

Fig. 52.

^MnnjTE Characteristics of Handwriting Lines.

(After Frazer.)

The

by two methods, the statistical and the microscopical. First, the measurements of the
latter are

studied

distance between certain letters in a given word, of the

ratio of heights to

breadths in certain

letters,

and

of the

^gulgx

slope of certain long strokes,

when

averaged,

ELEMENTS OF /IPPUED MICROSCOPY.

from which the deviations of the writer are
Composite photographs of signatures,
etc.,

yield values

rarely great.

are important aids in

place,

work

of this sort.

In the second

more minute

differences exist

which can be detected

only under the

compound microscope.
any long
lines

These are

of three

magnitudes.

First,

show

certain varia-

tions of direction

whose number and nature vary with

the writer.

Second,

much
from

finer fluctuations occur, visible

only under the

vertical

compound microscope,
side
to

in the shape of

deviations

side

and changes

in

width due to periodic changes in the pressure of the penpoint.

still

Third, on one or both margins of the line are

lateral serrations.

more minute

These

vertical

and

lateral variations are

are,

both illustrated in Fig. 52.

They
condiof

of course, influenced

more or

less

by paper and

writing utensils

and by the physical and mental

yet the size, position,

tion of the writer;

and number

the waves, swellings,

and notches are so related

to the

nervous organization of the writer, and differ so markedly

with different individuals, as to yield important aid in
the identification of handwriting.

REFERENCES.
Babcock,
J.

F.

The Medico-Legal Examination

other Stains, Hairs, and Fibres.

New

of Blood and York, 1894.

Phila-

Frazer, p.

Bibliotecs, or the

Study of Documents.

delphia, 1901.

Galton, F. Finger Prints. London, 1892. Galton, F. Finger-print Directories. London, 1895. RoscoE, H. E. Spectrum Analysis. London, 1870. Taylor, A. S. A Manual of Medical Jurisprudence. New York and Philadelphia, 1897. Wood, E. S., Withaus and Becker. Medical Jurisprudence, Forensic Medicine and Toxicology. New York, 1894.

CHAPTER

XI.

MICROCHEMISTRY.
I.

The Application
fields

of Microchemical Analysis.

In

few

have the

possibilities of the

microscope been

less fully realized

than in pure and applied chemistry.

science

which deals with the character and behavior

account

of chemical substances should surely take into

the characteristic crystals of which,

ditions,

under certain concomposed.

reactions

many such
trained
to

substances

are

Yet

chemists

study gross
ofteii

color

and

precipitates

have too

regarded the microscope as

strange

and impracticable instrument. and elaborate secondary

tests

Systems

of

recondite

have been

built

up, while the direct study of fundamental physical characters

has been neglected.

Although the toxicologist and

have used microchemical
tests

the medical microscopist

with such good results that in those special fields they

are

of

importance,

in
is

general the
slow;

adoption of these

methods by chemists

and only the develop-

ment

of

petrography, the

microscopic study of rock

sections,

has at

last,

during very recent years, called

general attention to their importance.

The study
to the

of microchemistry

is

by no means limited
141

examination of crystalline forms alone, as might

142

ELBMENTS op yiPPLIED MICROSCOPY.

its

be supposed from a perusal of

published by Harting in 1866.
others have since

earliest

exposition,

Lehmann, Behrens, and

shown

that

many dynamic

properties,

melting and volatilization for example, as well as the

typical

chemical reactions,

may

often be observed to

great

advantage

under the microscope.

The
results

use

of

minute quantities of material (one-tenth of a milligram

often sufficing), the rapidity with

which

may

be

obtained, and the simplicity and compactness of the ap-

paratus needed are general advantages of the microchemical

method, which apply in

all

manner

of analyses.

On

the whole, few chemists

who

take the trouble to familiarize

fail to find

themselves with the use of the microscope wiU

it

at times a valuable aid in their

work.

In the study
it

of certain closely related organic

at
if

compounds amount
^It

furnishes

once information which can otherwise be attained,

at aU, only
2.

by the expense

of a vast

The Study
student
of

of Typical Crystals.

of labor.

will

be well

for

the
first

microchemistry

to

familiarize

himself

with the most important characteristics used in the

identification of crystals,

by the examination

of certain

typical forms.

The

actual process of deposition should

in

be studied and those differences observed which occur

the formation of
as,

crystals

under

different

conditions,
solutions,

for instance,

from alcoholic and aqueous

and by slow spontaneous drying on the one hand and

rapid evaporation by heat on the other.
in a mother-liquor of the

Crystals formed
their

same composition show

and the

characteristics with special distinctness.

The

linear dimensions

relative proportions of

M/CROCHEMISTRY.

143

the various faces of a crystal vaiy widely with the supply

of material in the solution

from which they were deposited.

The
ever,

interfacial angles

between corresponding faces and

are,

the facial angles

between corresponding edges

how-'

always constant for a given substance.

as the

This law,

known

Law

of Steno, forms the basis for determina-

tive crystallography,

and makes

it

possible to distinguish

the crystals of such

compounds

as are not isomorphous.

Besides the general crystalline form, there are numerous

special points to

be noted in the study of a

crystal.

Its

habitus (the size and proportions due to the conditions

attending
its

its

deposition) should be noticed, as well as

refractive index, indicated

color

and the approximate

by the

definition of the edges either in air or in

some

denser mounting medium.

light,

The

effect

upon polarized
is

which

will

be discussed in Chapter XII,

often

of great importance.

Various peculiar phenomena

may

appear, such as

hemimorphism

(differences in the opposite

ends of a crystal), twinning (the production of double

crystals),

skeleton

crystals

(parallel

or

symmetrical

aggregates of smaller crystals), trichites (hair-like crystals,

often

more or

less

twisted),

and

sphseiulites

(radially

fibrous spherical bodies).

Sodium
and

chloride crystallized

rapidly

from a solution thickened with mucilage exhibits

skeleton
crystals;

beautiful

long, curved trichites

appear in a mixture of the chlorides of chromium and

mercury.

The change
salts furnishes

in crystalline

form of certain compound

an interesting subject for study under the

microscope.

If ferrous chloride is allowed to crystallize


144

ELEMENTS OF

/IP PLIED

MICROSCOPY.
large, tabular,
If,

from a warm,

aqueous solution,

monoslide

symmetric (rhombic) crystals form. be quickly heated by

now, the

the application of a small flame,

the large plates fall tp pieces, the

and minute

crystals
(Fig.

of

anhydrous
cooling, the

salt

appear in their place

53).

On

small crystals take

up water and run

together in their original form.

Fig. S3.

Crystals of Hydrous and Anhydrous Ferrous Chloride. (After Lehmann.)

Behrens and other authors give a large number of

specific

microchemical

tests,

some

of

which the student

should
of

make

in order to gain practice in the recognition

typical

crystalline forms.

Aluminium, for example,

may

be

easily detected in a solution

by evaporating with
Large,

a small drop of sulphuric acid, dissolving the residue in

water,

and adding a grain

of cassium chloride.

colorless, isometric

octahedra of caesium alum are pro-

duced,
ent,

or, if

more than

1% of

aluminium sulphate be

pres-

rectangular dendrites sprout from the caesium salt

(Fig. 54).

Calcium

is

best tested for

by precipitation with

sul-


MICROCHEMISTR Y.
145

phuric acid, which in concentrated solution throws out

short orthorhombic crystals of

anhydrous calcium

sul-

FiG. 54.

Crystals of C-esidm Alum.

dilute

(After

Lehmann.)

phate.

From

acid

solutions

slender monoclinic
crystallize out

prisms of the

compound (CaS04+ 2H2O)

Fig. 55.

Crystals of Calcium Sulphate.

(After

Lehmann.)

(Fig. 55),

showing numerous double twins, as at C,

masses of minute needle-

and

in presence of strong acids,

like crystals, as at .4.

146
3.

ELEMENTS OF APPLIED MICROSCOPY.

The Detection
earliest

of Quinin.

^It

has been noted that

one of the

and most

profitable applications of

microchemistry has been

made

in the study of

drugs

and poisons;
extent
path's

and the

toxicologist

depends

to

a great

upon various
method
to

specific

microchemical

tests.

Hera-

for the detection of quinin, applied par-

ticularly

the

examination of the urine of patients

under quinin treatment,

It

may

be studied as an example.
crystals

depends on the formation of characteristic

of quinin

of the iodosulphate

on the addition

of tinc-

ture of iodin to an alcoholic solution of the drug.

practice, the urine to

In

be analyzed

is

neutralized
is

and shaken

out with ether.

The

ethereal solution

evaporated and

the residue dissolved in a mixture of 12 cc. of glacial

acetic acid

and 4

cc. of

95%

alcohol, to

which

drops

of dilute sulphuric acid (i

gram

of strong sulphuric acid

with 9 grams of water) have been added.

very minute

drop of tincture of iodin added to

duces,
if

this solution first prospot,

quinin be present, a cinnamon-yellow

due

to the reaction of the iodin

little

and qumin.

Next

the

alcohol separates in

drops, driving the fluid away

Finally,

from the center of the preparation.

liquid flows

the acid

back again, and

thin, greenish plates of the

iodosulphate appear, often arranged in beautiful rosette

forms.

The

crystals

produce strong polarizing

eSects,

and turn brownish red when heated.

3.

The Separation

of Related Organic Substances.

The most

important aid which the microscope can

offer

to the chemist lies in the

domain

of

organic analysis^

Lehmann even

goes so far as to say that " Crystallography

.'aa

MICROCHEMIS TRY.
is

147

to the organic laboratory

what spectrum analysis

is

is

to

the

inorganic."

When

it

necessary to

separate

two closely related isomers, to determine the identity

of

two substances prepared by different methods, to

of elementary composition

distinguish differences

from

those due to allotropism, or to detect minute admixtures

of

an unpurity,

it

frequently occurs that purely chemical

methods are tedious and complex, while microscopical

examination furnishes a rapid and easy solution.
general the

In
the;

method

of study

consists

in allowing

substance in question to crystallize in contact with a

of

body

This

known composition, with which it is to be compared.^ may be done in three ways by adding to one

substance, in a melted condition or in solution, a crystal

of the other,

by preparing a mixture
from films

of
to

both in a liquid
cool,

condition

and allowing the mixture

and

bestj

perhaps,

by

crystallizing

of

the

two sub-

stances brought just in contact with each other.

When
till

solid substances are to

is

be compared, a small

portion of one
it

placed under a cover-slip and heated

melts, the

amount

of material being so small as

not to reach the edge of the cover.

grain of the second

substance
in turn

is

then placed just outside the cover-slip, and

first

melted so as to flow under and join the

along
(In-

a line which can be observed under the microscope.

cidentally
it

may be

noted that by heating two bodies

is

simultaneously under the microscope a good measure

obtained of slight differences in melting-points.

the
similar
crystals

Thus
cool

of

dinitrobenzol

and

dinitrotoluol

may be
and

easily distinguished.)

As two substances

recrystaUize, the

phenomena

along the contact zone


148

ELEMENTS OF APPLIED MICROSCOPY.

and
striking.

are diverse

In some cases the mixture

of

the two materials remains Hquid longer

han

the pure

substances, on each side, forming a clear area in which,

later,

an amorphous precipitate

is

deposited.

The

chlo-

rides of iodin
(Fig.
56).

and

silver exhibit this case to perfection

Or a

crystalline

precipitate

different

from

a.

b.

c.

Fig. 56.

Contact Zone between Chlorides of Silver and Iodin.

(After

Lehmann.)

either of the pure substances

may form

in the contact

zone,

as

with silver and potassium iodides.

Or

the

crystals

may grow

across the line unchanged, forming

a single homogeneous mass.

This occurs with

identical

bodies and with isomorphous substances like dibrom-

benzol and dichlorbenzol.

Or

the crystals

may grow
of the

across the contact zone, but with spaces between them,

due

to the presence of impurities in

one or other

substances.

Usually the habitus of the crystals in the

will

impure

substance

be

somewhat

changed.

Pur&.

dinitrotoluol

and the same body containing

phenomenon.

paranitro-

toluol exhibit this

These four cases by no

means exhaust

the possible conditions of the contact zone,

but they serve as types of some of the more striking ones,

MICROCHEMISTRY.
and

149

may

suggest

how

the microscope serves to test the

identity
5.
sis.

and the purity of organic bodies.

microchemistry

The Methods of Systematic Microchemical Analyhave seen

that
is

^We

much

wider term than crystallography, and covers the study

of change, of state
crystalline

and
In

of chemical reactions as well as

form.

this

broad sense,

specific

micro-

chemical tests

have been suggested by Behrens and

others for the detection of the elements

and the various

acid radicles.

Recently Hinrichs has prepared a com-

plete outline for qualitative analysis

under the microscope.

into general use is

Whether such a scheme

doubtful, although
of time
it

will ever

come

promises results with a

of

minimum

and material; but the study

two typical basic

and acid groups may prove suggestive

to the student.

A microburner yielding a flame not more

of

than a quarter

an inch in height must be provided for melting the

it

substances examined, and

is

convenient to have the

lamp so small that

the microscope
if

it

may

be placed under the stage of

desired.

For ordinary work micro-

scopic slides are used, a drop of the substance to be tested

being placed thereon with a fine glass stirring-rod.

reagent
is

The
to

added in minute quantity and

is

made

mix

with the
limation

first

by means
two

of a dry stirring-rod.

For sub-

tests,

I -inch

watch-glasses are used, one

inverted over the other, with the

substance to be extest

amined

in the lower,

and the appropriate

reagent

borne on the concave surface of the upper, glass.

6.

The Determination

of the Metallic Elements.

^Ac-

cording to the scheme prepared by Hinrichs, solutions of

15

ELEMENTS OF ylPPLIED MICROSCOPY.

first

the metallic elements are

tested

by the addition

of a

fragment of metallic zinc. If metallic crystals form,

silver,
is

either

copper, bismuth,
If

lead, tin,

cadmium, or thallium
zinc,

present.

a blackish coating appears upon the

gold, platinum, iridium, palladium, or

mercury

is indi-

cated.

Should

neither

phenomenon
tested for

follow, the other

groups of elements
nitric acid,

may be

by the

addition of

magnesium ribbon, ammonium

chloride,

and
are

magnesium and ammonium-phosphate

here,

solution;
first

we

however, concerned only with the

group of

metals, as typical of the general

If the solution contains

method

of analysis.

one of the metals precipitated

tested with several reagents,

by

zinc,

drops of

it

must be

each of which points out certain of the elements. a grain of potassium chloride
lead,
is

Thus
silver,

added, and with

is

and thallium a

characteristic reaction

obtained.
precipi-

Silver salts

produce an immediate white curdy

tate of silver chloride, lead salts, crystals of lead chloride,

and

thallium

compounds, very

minute

characteristic

crystals of thallium chloride.

tests

In each case confirmatory

silver,
first

must be applied.

With

for

example, the

amorphous

precipitate should be

separated by pour-

ing off the liquid and then dissolved in a drop of

ammo-

nium

hydrate.

The

colorless solution thus obtained on

warming and then cooling

silver chloride.

yields large cubical crystals of

original

If silver

be present, a drop of the

solution,

warmed and made

faintly acid with nitric acid,

large

produces on the addition of potassium bichromate

dark-red hexagonal plates and prisms of silver bichromate,

soluble both in nitric acid

and ammonia.

grain of

MICROCHEMIS TRY.
metallic lead

151

added

to a solution of silver salts causes

the separation of metallic silver in the

form

of dendritic

skeleton crystals.

The

other elements are identified

by

similar specific

reactions.

Copper

is

at

once indicated by the color of

solution.

the reduced metal

and the

Bismuth

is

detected

by the large hexagonal plates

by warming with sulphuric

of

bismuth sulphate formed

acid

and again

cooling.

Tin

solutions, with

an acid reaction, on the addition of

crystals of

sodium iodide, produce characteristic

iodostannites

sodium
a white

and iodostannates.

Cadmium forms
the

precipitate with
7.

ammonium

hydrate.
of

The

Determination

Acid

Radicles.

In
it

studying the acid radicles in an

is first

unknown

solution,

necessary to determine the presence or absence of

organic acids.

This

may

be accomplished by mixing a

drop with a minute drop of a saturated solution of potas-

sium permanganate and adding concentrated sulphuric

acid.

In the presence of organic acids,

nitrites,
is

sul-

phites,

and hyposulphites, the pink solution

decolorized.
decoloriza-

The
tion

last three classes of

compounds produce

when

acetic acid

is

substituted for sulphuric acid,

while the organic bodies do not.

If organic acids

be absent, a drop of the solution

is

tested for volatilization with acetic acid

and sulphuric
placed on

acid successively.
the

drop of

silver nitrate

under side of the upper watch-glass serves to absorb

the' vapors

and record

their presence

by

precipitation.
in-

The

first

group of acids decomposed by acetic acid

'-"

cludes the nitrites, cyanides, carbonates, sulphites,

<-,,'-'c--''

hypo,--..

^^A

-^^'-'-.-^

^-^

152

ELEMENTS OF APPLIED MICROSCOPY.

by
sul-

group, unaffected by acetic acid but volatilized

phuric add, includes nitrates, acetates, chlorates, chlorides,

bromides, iodides, cyanoferrates, cyaniferrates, and

borates.

The

third

group of non-volatile adds com-

prises the sulphates, phosphates, arsenates,

(colorless

and

arsenites

solutions),

the

permanganates, bichromates,
solutions),

and chromates (colored

In each case

and the

silicates,

molybdates, and tvmgstates (precipitates).

specific

confirmatory tests

are
first

made
groups
silver

for each of the acids indicated.

In the two
twice,

the

volatilization

is

repeated

once with

nitrate
glass.

and once with lead acetate on the upper watchNitrates yield negative results with the
first

and
Ace-

octahedral crystals of lead nitrate with the second.

tates

show the long white prisms with rhombic ends

of
silver

characteristic

acetate,

and no
are

reaction with

the

second

reagent.

Chlorides

characterized

by

thick rhombic prisms of lead chloride

crystals

and minute

cubical

of

silver

chloride.

In each case comparison

identify the

with

known substances makes it possible to compound present without serious difficulty.

REFERENCES.
Behrens, H. by J. W. CoHN, A. O.
Jtjdd.

Trans, of Microchemical Analysis. London, 1894. Tests and Reagents, Chemical and MicroscopicaL

a Manual

New

York, 1903.
F.

Hanausek, T.
HiNiLicHS, C. G.

Lehrbuch der technischen Mikroskopie.

Microchemical Analysis.
St.

Stuttgart, igoi.

Louis, 1904.
Phila-

Lehmann, O. Die Krystallanalyse. Leipsic, 1891. WoRMLEY, T. G. The Microchemistry of Poisons.

delphia, 1885.

CHAPTER

XII.

PETROGRAPHY AND METALLOGRAPHY.

I.

The Study

of

Rock

Sections.

The

determination

of minerals

can be more satisfactorily carried out by the

study of thin sections than by any other

microscopic

method, and the microscope occupies almost as important

a position in the geological laboratory as in that of the
biologist.

Frequently

optical

characteristics

are

the

only ones which can be relied

on

for the identification of

a given mineral, and almost always they are

more

easily

made

out than the chemical composition.

The

presence

and character of minute impurities are detected with

facility

under the microscope, and much information

may

be obtained with regard to the physical conditions which

attended the genesis of rock formations.

The

study of

the minute structure of fossils forms a subordinate but

attractive

branch of the general subject.

it

For examination with the microscope,

to prepare sections of rocks so thin as to

is

necessary

be transparent;

and

this is

a task of some
is

little

difficulty.

When

the

necessary apparatus
lathe,

at hand, slices are first cut with

using for a saw

tin

a thin disc of iron, copper, or

sheet

charged along the edge with diamond-dust

It
is,

or fine

emery.

however,

generally

possible
iS3

to

154

ELEMENTS OF /tPPLlED MICROSCOPY.

detach with a

hammer

a chip of suitible form without

using a special section-cutter.

The

;hip or section

is

then ground

down on an emery-wheel or by hand,

first

on an iron or copper plate covered with No. 120 emery

and water, then on a

glass plate with flour of emery.

is

The

slice

thus prepared

cemented

to a piece of glass

about one inch square.

as a cement, the

Canada balsam may be used specimen being pressed down into the
heated
till

warm

balsam, with care that bubbles are not included,

being then
the

the slide

balsam becomes

hard and firm.

section
is

After cementing, the free surface of the

until

ground down
has been

the

required- degree of the

glass serving as

transparency

attained,

a holder and support.

over 0.05

good section should not be

mm.

in thickness.
light,

Certain opaque minerals are observed by reflected

and

in

many

cases

it

is

convenient to examine mineral

in water for
in glycerin

powders

directly.

They may be mounted

the study of the form of the particles,

and

or

some

similar highly refractive substance for internal

structure.

The
scope.

color

and form

of the crystalline or amorphous

first

constituents of a rock are

observed under the microrelief indicate differ-

Appearances of high or low

of
refraction.

ences

The

presence

of

cleavage

liii:o

and

the occurrence of inclusions of gaseous, liquid, or

glassy character should be noted.

The
due

types of crystals
interpret

present

may be made

out, care being

taken to

correctly the peculiar appearances

to the plane in
to
lie.

which the section studied may happen

Even

the

PETROGRAPHY AND METALLOGRAPHY.

angle of the crystals can often be

ISS

measured with a micro-

scope provided with a revolving circular stage, such as

should be used for

all

petrographical work.

The
is

inter-

section of cross-hairs placed in the eyepiece

to coincide

adjusted

with the vertex of an angle, and one of the

is

cross-hairs

made
and

to coincide with one side of the crystal

lies

the stage

is

then rotated until the adjacent side

the angle of rotation

along

the cross-hair

measured on the

graduated edge of the stage.

definite outlines,

Incomplete crystals without

corroded crystals affected by the molten

strained crystals,

magma, broken or
of incipient crystals

and various types

Finally no pe-

may be made
is

out.

trographic

examination

complete without the study

of polarizing properties.
it

Before going further, therefore,

its

is

necessary to describe the micropolariscope and

appHcation.
2.

The Micropolariscope.
of

Ordinary
in
all

light

is

made

up

vibrations

of

ether

possible

directions

which he in a plane at right angles to the direction of

transmission of the light-ray.
particle

The path

of

an individual

of ether in ordinary light

would therefore be

constantly changing.
to the

Certain substances offer resistance

its

passage of such a light-ray, and confine

vibra-

tions to

two planes

at right angles to

each other.

Light

which, like that of the two resultant rays, vibrates only

in

ong plane,

is

known

as plane polarized light.

at

The
right

two rays produced, with vibrations in planes

angles to each other, obey different laws of refraction.

Thick layers

of

doubly refracting bodies

like calcite cause

so wide a separation of the

two rays as

to

produce two


156

ELEMENTS OF APPLIED MICROSCOPY.

re-

images of an object viewed through them (double

fraction).

The
sists,
is

polariscope

is

an instrument for detecting

refraction of bodies,

the

polarizing

power or double
cutting a

and

con-

in essence, of a pair of Nicol prisms, each of which

made by

rhomb

of calcite diagonally and

cementing the halves together again, with a layer of

Fig. 57.

Diagram of a Nicol Prism.

(After Clark.)

Canada balsam between them.

(Fig. 57)

Through such a prism

light

one of the rays of plane polarized

(EFGH),
strongly
it

known
the

as the extraordinary ray, passes unchanged, while

other

(EFKL),
first

the

ordinary ray,

is

so

refracted in the

half of the prism that

at

meets
totally
re-

the layer of
reflected

balsam

such an angle as to be
surface,

from the balsam

and

is

thus

moved.

Nicol prism

is,

then, a device for removing

PETROGR/IPHY AND MET/1LLOGR/1PHY.

I

157

the ordinary ray

from plane polarized

light

and allowing

only the extraordinary ray to pass.

For studying polarization phenomena, the microscope

is

provided with two Nicol prisms, properly mounted,

one placed below the stage and

known

as the polarizer,

the other inserted, as a rule, in the tube of the microscope

above the objective, and

or both
jof

known

as the analyzer.

One

the Nicols

and the stage

of the microscope

are arranged to be rotated at will.

The

polarizer breaks

up the
Ught,

light

from the mirror into two rays of plane polarized

of these

and

suppresses the ordinary ray.

The
the

remaining
stage

extraordinary

ray

passes

up

through

and objective

of the microscope to the analyzer.

When

the analyzer occupies

two positions
at 180

relative to the

polarizer,

one in which the oblique cut surfaces of the

two are

parallel,

and another

from

this, it is as if

they were continuous,

passes freely.

and the ray

of plane polarized light

At the intermediate points, 90 frorn the no Hght passes

at
all,

parallel position,

since the extraor-

dinary ray bears to the second prism the

same
is

relation

which the ordinary ray does to the

suppressed.
into

first,

and

therefore

At intermediate points the ray

is

broken up

two components, with vibrations

at right angles to

each other, and one of these components passes the

analyzer,

producing a sort of twilight, the intensity of

is

which increases as the parallel position

If,

approached.

now, the polarizer and analyzer be crossed, placed,

is,

that

at 90

from the

parallel position,

no

light passes

and the

field of the

microscope remains dark.

01

If crystals

of sodium chloride

some other

salt

crystaUizing in the

15^

ELEMENTS OF APPLIED MICROSCOPY.

mounted on a
slide
still

isometric system be

and placed on
remains dark.

the stage of the microscope, the stage

Conditions are unchanged, since

like glass

amorphous substances
system produce

and

crystals of the isometric

effects.

no polarizing

When

calcium-carbonate crystals,

or those of any other substance of the last five systems,

are examined with crossed Nicols, the crystals
visible as shining bodies
cise

become
exer-~

on a black ground.

They

a polarizing action of their

own upon

the plane polarit

ized light which passes through them, breaking

into

up

two rays polarized

at right angles to

each other and

is

not coincident with the plane in which the light

sup-

pressed

by

the

analyzer.

Thus,

when viewed with

become
bodies do

crossed Nicols, polarizing or anisotropic bodies

luminous, while non-polarizing or isotropic

not.

One

other

phenomenon must be noted

by

in the

most

rudimentary account of the polariscope, the production

of color effects
If a plate of

thin plates of anisotropic substances.

of the micro-

mica be placed upon the stage

scope, the entering ray of plane polarized light,

which we

may

call

A,

is

resolved into two rays with vibrations at

right angles to each other,

A' and B'.

Of

these, the or-

dinary ray B' at right angles to

A and A' is most refracted,

retarded by an amount

passes through a greater distance in the mica than does

the extraordinary ray,

and

is

depending on the thickness of the plate and the strength

of double refraction of the substance.

In the analyzer

each of the two rays A' and B'

is split

up again -into two

components

at right angles to

each other, A' a and A'h,

PETROGRAPHY AND METALLOGRAPHY.

1^9

B'a and B'h. A' a and B'a are parallel to the plane of the
original

illuminating ray A,
lie

and are suppressed.

A'b

and B'h
freely.

in the plane at right angles to this

and pass

As they
due

unite, however, their wavesiare, in differ-

ent phases,
the

to the retardation of B' in passing through

mica

plate,

and

if

the

amount
will

of this retardation be

right, interference

phenomena

be

set

up and very

beautiful color effects produced.

The

particular color

varies with the thickness of the plate

examined; but at

the two opposite positions of the analyzer at

light passes, the colors

which most

which appear are complementary.

illustration, in

Using the notation adopted above for

case A'a and B'a,
since the
in the other,

one

A'b and B'h, pass; and,

sum

of all these four rays originally produced

white

light, it is

obvious that the complementary relation

must
3.

exist.

The

Identification of Minerals.

Besides the general

it

microscopical appearance of crystals noted above,

is

possible with the. polariscope to study one character of

perhaps more practical importance than any other, the

optical structure as indicated
light.

by

its

effect

on polarized
or isometric

We

have seen that crystals of the

first

system are isotropic.

All other crystals are anisotropic,

but in those belonging to the tetragonal and hexagonal

systems there
crystals is
is

one axis about which the structure of the

this axis light passes

homogeneous, and along

unaffected.

Such

crystals

are

called

uniaxial,

while

crystals of the orthorhombic, monoclinic,

and

triclinic sys-

tems possess two such axes, and are called

biaxial.
it

Examination with crossed Nicols makes

possible at

i6o

ELEMENTS OF /tPPUED MICROSCOPY.

once to distinguish isometric crystals from those of other

systems.

Good

crystals to

compare

in this respect are

garnet and gypsum, the former being isotropic, the latter

anisotropic.

The same

minerals furnish an

instructive

contrast in refractive index, the former showing high

and

the latter low .reHef

Tetragonal, hexagonal, and orthorhombic crystals

may
tri-

be separated from those

clinic types

of

the

monoclinic

and

by the

fact that in

them the

directions of

vibration (sometimes called axes of elasticity) are parallel to

the crystal axes of the crystal.

The

position of

these vibration directions can be determined

by noting
(position

the position in which the crystal

of extinction)

becomes dark
it

with crossed Nicols, for

is

then that

the vibration directions coincide with the planes of the

Nicols.

cross-wire placed in the eyepiece so as to

coincide with the plane of vibration of the analyzer under

these conditions coincides also with the axis of elasticity
of

the

crystal.

By now removing

the

analyzer,

the

cleavage lines and boundaries of the crystal

may

be

seen and the stage rotated so that the cross-wire corre-

sponds with them. produce

this effect as
if

The

angle of rotation required to

of

measured on the graduated edge

the stage,

o or 90, on all the crystals examined, indi-

cates a tetragonal, hexagonal, or orthorhombic crystal.

Such substances are said

or to be symmetrical.
condition, while in

to

show no

extinction- angles
illustrates this
all

Quartz (hexagonal)
(monoclinic)
angles.

Gypsum
extinction

planes but
like

one

show

large

This,

other

monoclinic minerals, exhibits in one plane phenomena


PETROGRAPHY AND METALLOGRAPHY.
similar to those characteristic of hexagonal crystals,

i6i

and

shows no extinction angle.

When

certain sections of anisotropic crystals are

exam-

ined with convergent polarized hght obtained by a con-

verging lens fitted over the polarizer, interference figures

are produced, consisting of dark or colored rings
crosses.
It is impossible to enter here

and

into the

optical

principles difference

which condition these phenomena;

but the

between the circular figures produced by uni-

axial crystals,

and the

elliptical

appearances characteristic

Fig. 58.

Inteeierence Figures.

(After Luquer.)

of biaxial crystals (Fig. 58), are easily apparent.

With

some

sections

which show only an

if

indistinct

bar instead

of a clear figure,
it

the stage be rotated, the bar follows

and remains

straight, in the case of uniaxial crystals,

it

while with biaxial crystals

tion

moves

in the opposite direcwill again

and becomes curved.

Quartz and gypsum

serve for comparison, the former being uniaxial, the latter

biaxial.

Two more

characters remain to be noted, absorption

i62

ELEMENTS OF APPLIED MICROSCOPY.

Absorption
is

and pleochroism.
all

the

power

of absorbing

or part of the light which vibrates in certain planes,

similar property of

and pleochroism the

section be

removing the
If

rays of certain colors in particular planes.

a rock

examined with

parallel rays of polarized light

and the
tion will

stage be rotated, minerals

which exhibit absorpfrom the

show a change

in intensity of light

usual shade almost to black, while pleochroic minerals

show a change

in color, or at least in shade.

sufficient to identify

We
six

have

now data

any one

of the

crystallographic

systems, whose

optical

characters

may
gent

be briefly summarized as follows: Isometric system:

isotropic, exhibiting
light.

no interference

figures in converuniaxial,
inter-

Tetragonal system:
symmetrical,
sections

anisotropic,

extinction

giving

uniaxial

ference figures, often with rectangular cleavage.

Hex-

agonal system: anisotropic, uniaxial, extinction symmetrical,

sections giving uniaxial interference figures, three-

or six-sided, or
of 60.

show cleavage

lines intersecting at angle

Orthorhombic system: anisotropic,

symmetrical in
all

biaxial,

exsys-

tinction

sections.

Monoclinic

tem:

anisotropic, biaxial, extinction symmetrical in one

zone, while other sections

clinic
all

show

extinction angles.

Tri-

system:

anisotropic, biaxial, extinction angles in

sections.

By

application
of

of

these

criteria,

comoptical

bined

with

observation

the

more obvious
it is

characters, color, aggregation, etc.,

possible to identify

any
4.

of the

commoner minerals with

ease
of

and

certainty.

The Structure and Behavior

Alloys.

Metal-

lography, or the study of the structure of metals with the

PETROGRAPHY AND METALLOGRAPHY.

microscope, owes
its

163

beginnings to Dr. Early of Sheffield

Its

and Professor Martens of Charlottenburg.

aim

is

to

study the minute structure of metals with the microscope,

not, as in the case of minerals,

by grinding off thin sections,

light.

but by examining polished surfaces by reflected

Its

importance

steel as to

now so great in demand some attention

is

the testing of iron

in the

and

most elementary
it is

survey of the field of microscopy; and since

various alloys that metallography
is

with the

principally concerned,

certain general characteristics of these bodies

must

first

be briefly discussed.

Many alloys may,

and copper
is

for practical purposes, be considered

as solutions of one metal in another;

the alloy of silver

If a

a good example of this type.

molten

alloy containing

more than 72%

is

of silver

be gradually

cooled, a point

reached at which the rate of cooling

silver

becomes retarded and

greater the
ture at

begins to separate.
is

The
when
silver

amount

of silver the higher

the tempera-

which

this process begins.

After a time,
the
still

the temperature falls to 770

will

C,

molten

be found to have fallen to just J2%; a second retardation in the cooling is now apparent accompanied by the
soHdification of the entire alloy.
If

more than 28%

first

of

copper be present, on the other hand, the

is

retardation

accomplished by a deposition of copper, which con-

tinues until the excess has all been solidified,

at

which occurs

a temperature of 770 C.

The remainder then soHdifies

containing
originally

entire.
silver
tion,

molten alloy

72%

of

and

28%

of copper

shows only one point of retarda-

remaining molten above 770 and becoming solid

164

ELEMENTS OF

y4P PLIED

MICROSCOPY.

below that temperature.

points
is

The

relation of these various

made more

clear in Fig. 59, in

which the outer

limits of the oblique lines represent the fusing-points of

pure

silver

and copper.

saver 100

PETROGRAPHY AND METALLOGRAPHY.

S.

165

Metallography

of

Steel.

Steel

is

practically

an

alloy of ferric carbide

and

iron,

and the phenomena attend-

ing
the

its

cooling from a high temperature are essentially

just considered.

same as those we have

The com-

pound corresponding
FegC, or

to the eutectic alloy contains

12%

0.8%

of pure carbon.

This substance

is

known
concar-

as pearlyte,

and

steel of the right

composition on gradual

cooling from a fused condition

becomes

entirely
If

verted into

it

at a

temperature of 670 C.
carbide
if

more

bon be present,

ferric

is

separated before this

temperature be reached;

iron, that

metal

is

segregated.

The
up

carbide

is

known

to microscopists as cementite, the

pure iron as
of ferrite,

ferrite.

Very low carbon

steels are

made
of

with here and there particles of pearlyte.

of pearlyte increases with

The proportion
carbon up
cementite.
If steel

an increase

to

0.8%,

after that point being

mixed with

be suddenly cooled from a high temperature,

a

above that at which segregation- begins,

new comto this sub-

pound
stance

is

formed known as martensite, and

due the intense hardness of
at
steel

is

thus quenched.

Below the point

separate,

which

ferrite or

cementite begins to

sudden cooling produces a mixture of these

bodies with martensite.

If a

smooth surface of steel be examined under the micro-

scope the presence and the general proportions of these

various

compounds may be

readily
size,

made
is

out.

The
very

specimen, of some* convenient

to

say from half an inch

first

an inch square and

half an inch thick,

tace with

carefully polished

upon one

emery and with

^66
rouge.

ELEMENTS OF APPLIED MICROSCOPY.

by rubbing the metal on a layer of parchment stretched over smooth wood and covered with moist rouge to which a little

The

final polishings are carried out

ammonia-water has been added.

the softer parts are
structures stand out in
finally

In this latter process

somewhat eroded and the harder

low
relief.

The specimen when

prepared

is

examined with an ordinary micro-

scope provided only with some apparatus for illuminating

with reflected

light.

This

may

consist merely of a con-

denser or reflector which throws light obHquely on the

stage, or better, of
fitting into the

a plane reflector or right-angled prism

tube of the microscope above the objective.

its

This

is

perforated at

center to allow passage of rays

its

upward from
reflects light

the object, while the remainder of

surface
the

from without

vertically

downward on

specimen.

When examined
pearlyte
is

in this fashion with the high objective,

seen to be

made up

of very

minute

crystals of

two substances arranged alternately;

teristic

this is the charac-

structure of all eutectic alloys,

and

indicates that

in spite of their constancy of composition these bodies are

merely peculiarly intimate mixtures of the two constituent substances.

Cementite,

if

present,

appears in
it

large

whitish

masses;
relief.

being harder than pearlyte,

stands out in

Ferrite,

on the other hand,

is

the

softest material in steel;

of free

and in alloys with more than 88% iron pearlyte areas appear standing out from a
ferrite.

background of
of

Fig. 60 illustrates the structure

as

mixed

pearl)'te

and cementite

seen
it

under the

microscope.

If

martensite be present

appears as a


PETROGRAPHY AND METALLOGRAPHY.
homogeneous substance made up
easily distinguished

167

of

ciystalline needles

from any other constituents

of

of steel. deter-

The

exact

amount

carbon in

steel

must be

FiG. 60.

Microscopic Appearance op Steel with 1.5% Carbon (Pearlyte and Cementite), (After Sauveur.)

mined by chemical methods;

mineral forms

but the presence of the

we have

discussed and the

manner

in

which they are associated can only be ascertained under

the microscope.

Furthermore, the crystals present

arranged,

may

be

abnormally

being

perhaps

not

closely

packed or so large as to produce incipient cleavage planes.

68

ELEMENTS OF APPLIED MICROSCOPY.

All these factors are of great importance in determining

the hardness of steel;
it

is

therefore natural that the

microscope should have become indispensable in

laboratory of the iron and steel expert.
Its

the

practical

apphcation to the study of other alloys promises extensive

development in the future.

REFERENCES.
Clark, C. H.
1894.

Practical

Methods

in

Microscopy.

Boston,

LuQUER, L. M. Minerals in Rock Sections. New York, 1898. Zerkel, F. Lehrbuch der Petrographie. Leipsic, 1893. RosENBUCH, H. Iddings, J. P. (translator). Microscopical Physiography of the Rock -making Minerals. New York,
1893.

HiORNS, A. H. Metallography. London, 1902. Sauveur, a. The Constitution of Steel considered as an Alloy of Iron and Carbon. Technology Quarterly, XI, 1898, 78.

INDEX.

PAGE

Abbe
" condenser
Aberration, chromatic
i9. 27,
12, 14, 35,

19

28 36 36
152

"

spherical

12, 14, 35,

Acetates, determination of

Achromatic objective (see Objective).

Acid haematin " radicles, detection of
Acids, detection of

134
151
15

Adjustable objective

32

Adjustments (see Microscope, adjustments).

Adulterants of drugs (see Drugs).

" Agave

" foods

(see Foods).

89
119

Agglutinitis.

Air-bubbles

30
49
144
44, 45

Albumin fixative Aluminium, test for

Alcohol, as a fixing agent

Alhazen
Allotropism
Alloys, behavior of

10 147 162
,

"

nature of

163 91

Alpaca
Alterations in manuscripts

138
15. 17

Amici

Ammonium-magnesium phosphate
Anabcena in water

in urine

107 123

169

I70

INDEX.
PAGE

Anaemia Analyzer Angiosperms, fibres of Angle of prisms Angular aperture Animal hairs (see Wool).
Anisotropic bodies

'

"3
'57 loi
S

17

158
135 17
17

Anthropometry
Aperture, angular

"

.numerical

Aplanatic lenses fsee Objective).

Arabian microscopy

lo

Aristophanes

Armati
Arrowroot
Asbestos
Asterionella in water

9 lo

66
81

124

Axial illumination
Axis, principal

26 6
117 116 10

Bacillus diphtheria

" Bacon Balsam

tuberculosis

(see

Mounting Media, balsam).

84
142, 149

Bast-fibre

Behrens Bertillon
Biaxial crystals

135

159

Biconvex lens
Birch fibres

(see

Convex Lens).
loi
tests for

Bismuth, microscopical

Ijl

Blood, examination of cells

log
II4,

"
'

"

for malarial parasites

'

spectroscopy of

133
Ill

Blood-cells

"
"

counting
in

,,2
107, 108

""ne

"
"

normal numbers
staining

U, Ul
jpn

Blood-smear, preparation
Blood-stains, detection of

j2g

INDEX.

171
PAGE

Blood-Stains, microscopical test

127

"

microspectroscopical

128

Bakmeria

88 92
ill

Bombyx
Basophiles.

Brownian movement
Buckwheat-starch
Burning-glass

37 78
9
tests for

Cadmium, microchemical
Csesium alum, crystals of

15

144
107
145

Calcium oxalate in urine

" "

sulphate crystals
test for

144
92
54, 55, 56
55,

Camel's hair

Camera Lucida " " construction of " " lighting with " " measurement with " " use.of Canada balsam (see Mounting Media, balsam).
, , , ,

56
55

54 56 86

Cannabis
Carbol-fuchsin

n6
167 91 105
cells).

Carbon in

steel

Cashmere
Cassavas (see Sago).
Cell theory
Cells (see

Mounting,

Cellular pathology

105
165, 166
15

Cementite

Chevalier
Chicory (see
CoflFee, adulterants).
.

Chlorides, detection of

152
143
45, 46

Chromatic aberration (see Aberration).

Chromium and mercury

Clearing agents
Coffee adulterants

chloride

"

microscopic structure

73 72

Color production by mica plate

158 100

Condenser (see ABBfi Condenser).

Coniferae, fibres

72

INDEX.
PAGE

Contact zone

'47

Continuous spectra

'3^

Convex "
"

lens, formation of

images
4t

"
o
1^3
15*

'

refraction

by
tests for

Copper, alloy with silver

,

microchemical

Corchorus
Corn-starch
59i
,

^7

^4
65
81

"
Cotton

microscopic structure
8r,

"

-fibre

82
82

"
" " " " "

"
" "

canal

chemical treatment

83 82 82 83 82
95
31, 32
32, 39

.composition
.length
.

" " "

mercerization

preparation

Cotton paper
Cover-glass, effect of

"

measurement of

Cover-glasses, cleaning

41
143
155

Crystal angles

"

"

.measurement

,Cryohydrate
Crystals (see Minerals).

164

.anisotropic
,

159.161,162
159 142 143
159. 160, 162
159. 162
,

biaxial

formation of
habitus

hexagonal
isotropic

microscopic study of

142
Ijg^ 160, 162 159^ 160, 162

monoclinic

orthorhombic
tetragonal
triclinic

I59_ 160, 162

159, 160, 162 I rg

, ,

uniaxial

Curvature of lens

6
'

"
Cytology

" the

field

j.
,oj

Dark-ground illumination

2-

INDEX.

ili
PAGE

Dehydration, in glycerin " with a Icohol ,

43
45 10
117

Descartes
Diagnosis of Diphtheria " " Tuberculosis

116 118

" "

" Typhoid Fever

(see
,

Diaphragm
Diatoms

Microscope, diaphragm).

use of

28
35. 124

Differential blood count

112
49i 5

"

staining

Dinitrobenzol
Dinitrotoluol

147 I47 117 137

Diphtheria, diagnosis of

Documents, study of
Double staining
Draw-tube, use of

50
20, 31, 32
II

Drebeel Drug adulterants

Drugs, microscopically examined for adulterants

69

69

Dry mounting

(see

Mounting Media,

air).

Early Ehrenberg
Eosin and hsematin, double staining
Eosinophiles
Epithelial cells in urine

163

106

50
Ill

109
Ill

Erythrocytes
Esparto-fibre

103

Eutectic alloy

164
2

Eye, structure

Eyepiece (see Microscope, lenses).

Eye-point
Eyes, care of

33

33 94
165
165, i6$

Fabrics, examination of Ferric carbide in steel

Ferrite

Fibres, analytical

key
81,

"

animal
of reagents

98 90
81

"

^ effect

174

INDEX.
PAGE

Fibres, kinds of

"
"
' '

.textile
,

80
8l
r

vegetable

wood-pulp

100
102

Fibrovascular bundles
Finger-print classification
' '

137 136
135

patterns

Finger-prints
Fir-fibre
.'.

100

Fixative,

Mayer's albumin

49 44 44 84
85
44,

Fixing

"

agents

Flax " -fibre

Flemming's Mixture
Focussing
Focus, principal

29

6 69
of.

Food adulterants
Forgery, detection

138
15

Fraunhofer
"
lines

132

Freezing microtome

49
11

Galileo

Galton Germ theory

of disease

135 106

Glycerin (see Mounting Media, glycerin).

Glycerin jelly (see Mounting Media, glycerin

jelly).

Goring
Gossypium

15

81
(see Trichina).

Government inspection of meat

Gower's

solution
fibres

112

Gymnosperms,

100
l6i

Gypsum
Haemalum, Mayer's
HtBfnavKBba
,,,
eosin, double staining

50
114,

Hsematin and

50
li j
j

Hsematin, spectrum of

Haeraocytometer

j2

Haemoglobin, spectra of

j,^

INDEX.

175
PAGE

Handwriting

138
-lines,

"
"

microscopic appearance

of.

140 139
142
143

statistical

study

of.

Harting Hemimorphism
Hemlock-fibre

100

Hemp
"
-fibre
,

86
86

" "

Manila (see Manila Hemp).

Sisal (see Sisal

Hemp).
135 105

Herschel Hertwig ...

HiNRICHS Homogeneous immersion

149 16

HooKE
Huyghenian ocular
Illuminating power

104
22

35

Illumination (see Lighting).

Image, construction of

7,8
i 7, 7,

"
' '

definition of

formation of by object inside principal focus

formation of by object outside principal focus
real

"

" "
'

7,

8
i

retinal
retinal,

'

formation of

3 of 2

" "

retinal, interpretation

.virtual

7,9
of,

Images, formation in
' ' ,

formation

compound microscope by convex lens

II, 13

Imbedding in celloidin " " paraffin

Immersion, objective (see Objective).
Infusoria in water (see Water, Drinking-).

49 47

Ink, examination of Interference effects

137

158

Interpretation of appearances

36
2

"

" retinal image

lodin chloride, crystals

148

lodo sulphate of quinin

Iron in steel

146
165

176

INDEX.
PAGE

Isomers, separation of
Isotropic bodies

'47 '5^

Janssen
Jute

II

87
-fibre

"

87

KiRCHER KOLLIKEK
Larch-fibre

10 105

100
tests for

Lead, microchemical

150
10, 104, 106

Leeuwenhoek Lehmann
Leishman's blood-stain
Lens, curvature of Leucocytes, various types Leucocytosis

142, 146
51, iii, 114

6
ill

113
25, 26, 27, 28

Lighting

Linen paper
Line spectra

95
132

Linum
Lister Loeffler's methylene blue Lymphocytes
Magnification, measurement of

84
15

117 112

57

Magnifying power
Malarial parasite

34
11+

Malpighi
Manila hemp

104 89
97
163 166
71, 122

"

paper

Martens
Martensite

Massachusetts State Board of Health

Mayer's albumin fixative haemalum Meat inspection (see Trichina).

' '

49

50

Mechanical stage
Mercerization
Metals, microchemical tests for

19

gj

150

INDEX.

177
PAGE

Metals, separation of in alloys

Metallic elements, determination of

163

Metallography " of

149 162
165

steel

Methylene blue
Microchemical analysis, apparatus for " " ofacids

117 149
151

"
" " "

"
" "
tests
,

"metals
possibilities of

149
F42

.systematic

149
144

Micrometer, measurement with " ocular

,

54
53 52 53
155, 157

"
"

stage

standardization of

Micropolariscope
Microscope, adjustments
'

20
20
23
10, 11, 12, 13

'

arm
cleaning

" "
' '

compound diaphragm
draw-tube
function of
history of

19

"

20
3 9 12

"
' '

"
"

HoOKE's
biology

in

104
105
II
20, 21

" "
' '

in
,

medicine and" sanitary science

Leeuwenhoek's
lenses

"
"
,

"

cleaning of

24
57
17

magnification of

"

mechanical parts
mirror
pillar

"

19 19

"
"

position of
qualifications of
setting

23

"
" "

up

34 23
9
19

simple

" "
"

.stage
stand
tube

17

20

178

INDEX.
PAGE ^2
'3
128
4^, 49

Microscopy, forensic
Microspectroscope

"

Microspectroscopy of blood-stains

Microtome
Minerals, identification
in rock sections

159-162
154

Minot-Blake microtome
Mohair
Motion under the microscope Mounting cells Mounting media, air
'

48
91

36
41) 42

41

" "

" " "

"

.balsam
.effect of
,

43

38
42
jelly

glycerin

"

43
38, 39

"
"

" "
"

refractive effects
requisites of

40 39 89 34
75

"

temporary

Musa Muscm
"

volitantes

Mustard, adulterants
,

microscopic structure of

74
22

Negative ocular
Neutrophiles Nicol prisms
Nitrates, detection of

112
156, 157

152
I

Normal

vision

Numerical aperture
Objective (see Microscope, lenses).

17

"
"

achromatic
aplanatic

14, 15

l6
15. 16

"

immersion

Oblique, illumination

27

Ocular (see Microscope, lenses).

Oil-drops
Olive-stones (see Pepper).

30
I^

Organic acids, detection of " substances, examination of

Overstaining
, . ,

146

jo

INDEX.

79

PAGE
Ovis
91

Oxyhsemoglobin, spectrum of
Oxyphiles

134 iii

Paper and paper making

95,

96 98 96 96
97
95

"
"
"

-fibres,

analytical

key

manufacture

"

raw materials stock, kinds and amounts

,

Papyrus
Parafiin

imbedding

Faranitrotoluol crystals

47 14S
95 106
165

Parchment

Pasteur
Pearlyte in steel

Pedesis
Penetration

37
35

Pepper, adulterants

7^

"

microscopic structure of

7^
153

Petrography

Phloem

(see Bast-fibre).

nper
Plano-convex lens (see Convex Lens).

76
155 15S

Plane polarized light

Pleochroism

Pliny
Polariscope

9
155, 156
structure of

"

156
157

Polarization with microscope

Polarized light
Polarizer

,....

155

Polarizing microscope
Poplar-fibres

157 I57
loi

Pork, examination for Trichina (see Trichina).

Potassium iodide crystals

Potato-starch

14S
59, 61

"
"
'
'

microscopy of

63 6

Principal axis

focus

6
5
,

Prisms, effect of angle

,

refraction

by

i8o

INDEX.
PAGB

Qualitative analysis, microchemical

149 I^l

Quartz
Quinin, test for

146
'

Ramie
"
-fibre

^^

88

Real image (see Image).

Reaumur
Reduced haemoglobin
Reflected light
,

95 134 28

Refraction by convex lens

4
3>

" "

" plane surfaces

4
5

" prisms
,

" "
Retina

laws of

toward normal
,

4
35
I,

Resolving power
Retinal image

2, 3

Retting
Rice-starch

84 66
154
153 153

Rock
"

sections, characteristics of

"

preparation of

"
Sago

"

study of

67
104
104
;

schleiden

Schwann
Scutching
Section cutting

84
46

"

"
"

" in pith Sedgwick-Rafter method

Selligues

" "

by hand " microtome..

46
.

47

46,47
122
IS

Seneca Serum reaction

"
Silk,
".
,

g I18
119

test for

typhoid fever

formation of
structure of

92
93

Silver, alloy

with copper
,

163

chloride crystals

148

"

iodide crystals

14S

INDEX.

l8i
PAtJE

Silver,

microchemical

tests for

150
89
143

Sisal

hemp

Skeleton crystals
Slides, cleaning

41
143 132

Sodium
"

chloride, crystals

Spectra of gases and solids

" organic bodies

133
131

Spectroscope
Sphserulites

143

Spherical aberration (see Aberration).

Spruce-fibre

100
115

Sputum, examination of
Staining

49
,

"
<<

blood-cells

iii

diphtheria bacilli
tubercle bacilli

L17
116
,

"
" " "

with haematin and eosin

5<->

Stage micrometer (see Micrometer).

(see Microscope, stage).

Starch, as index of food adulterants.

,

in baking

powder
,

69 62

"
"
"

commercial uses for

conversion into sugar

60
62
5^ 59 61

formula of
in chlorophyll in flour

"

"
" "
"

in grains

60
59< ^'

in potato

microscopy of
nature of
origin of

62
5^
58 61 59 60

"

"
" "

paste
,

preparation for market

prices of
refining of

"
"

"
State
Steel,
'
'

uses of

59 61

Starches of pea

and bean Board of Health of Massachusetts, work on food adulterants.

composition of

66
71
165

metallography of
b-vv

165

Steko,

of

143

82

INDEX.
PAGE 103
97
103

Slipa

'

Straw-board
Straw-fibre

SWAMMERDAM
Synura
Tapioca
in water
(see Sago).

IO4

124

Telescope, function of

3
tests for

Thallium, microscopical

150

Thoma

microtome

48
151

Tin, microscopical tests for

Toxicology
Tracheids
Trichina
Trichinosis (see Trichina)
Trichites
.'

146
100

120

143

Tube "

casts in urine

109

length, standard
bacilli

Tubercle

31 116

Tuberculosis, diagnosis of

116
15

TULLEY
Turntable

42
143

Twinning Typhoid bacilli, agglutination of " fever, diagnosis of

Ultra-violet light

119
1

18
21

Uniaxial crystals Urates in urine

159 107

Uric acid in urine

107
107
.

Urinary sediment Urine, examination of Uroglena in water

107

124
105 105

Van Beneden ViRCHOW

Virtual image (see Image).
Vision, normal

Walker
Water, microscopy of drinking

129
121
122

"

filtration

ol

INDEX.

183
PAGE 64

Wheat-starch

"

microscopic structure

64.

Whitney WiDAL reaction Wilder Wiley Wood pulp

'

97 119
135

60
97 ICO
9Si 97

".fibres

'

"

paper

Woodman.".
Wool, kinds and qualities " characteristics of
,

97

92
90
51, iii,

Wright,

blood-stain

114
137

Writing instruments

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Bernadou's Smokeless Powder. Nitro-cellulose, and Theory of the Cellulose i2mo, Molecule Svo, Bolton's Quantitative Analysis * Browning's Introduction to the Rarer Elements Svo, Svo, Brush and Penfield's Manual of Determinative Mineralogy Classen's Quantitative Chemical Analysis by Electrolysis. (Boltwood. ). Svo, i2mo, Cohn's Indicators and Test-papers
Tests

i2mo,

25 00 3 50 i 50
i

2 so

and Reagents
Course in Qualitative Chemical Analysis. (Schaefler.).
.

Svo,

Crafts's Short

.i2mo,

50 30 4 00 3 00 2 00 3 00 i so
i

Dolezalek's Theory of the Lead Accumulator CStorage Battery).

Ende.)
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(MerriU.)

(Von i2mo, i2mo,

Svo,

2
i

so

Duhem's Thermodynamics and Chemistry. (Burgess.) Eissler's Modern High Explosives (Prescott.) Effront's Enzymes and their Applications. Erdmann's Introduction to Chemical Preparations. (Eunlap.)
3

8vo, Svo,

25 4 00 4 00 3 00
i

i2mo,

25

Fletcher's Practical

Instruc;!;:::-::

r.-.:

^. :::;:::

::.

_-

--;
i2mo, morocco,
i

50

Works Analyses (Wells.) Fresenius's Manual of Qualitative Chemical Analysis. Manual of Qualitative Chemical Analysis. Part I. Descriptive.
Fowler's Sewage
2 vols

i2ino,

2 00

(Wells.) 8vo

System of Instruction in Quantitative Chemical Analysis.

Fuertes's

Water and Public Health Furman's Manual of Practical Assaying * Getman's Exercises in Physical Chemistry i2mo, i2mo, Gill's Gas and Fuel Analysis for Engineers (Woll.) i2mo, Grotenfelt's Principles of Modem Bairy Practice. 8vo, Hammarsten's Text-book of Physiological Chemistry. (Mandel.) i2mo, Helm's Principles of Mathematical Chemistry. (Morgan.) iCmo, mcrocco, Hering's Ready Reference lables (Conversion Factors) 8vo, Hind's Inorganic Chemistry * i2mo, Laboratory Manual for Students 8vo, Holleman's Text-book of Inorganic Chemistry. (Cooper.) Text-book of Orgahic Chemistry. (Walker and Mott.) 8vo, * i2mo* Laboratory Manual of Organic Chemistry. (Walker.)

8vo, 5 00 3 oo (Cohn.) 8vo, 12 50 i2mo, i 50 8vo, 3 00

2 00
i

25

2 00

4 00 i 50 2 50 3 00
73
2 50 2 50
1

00 25

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3 00
i

2 50

00 3 00
i

2 00
i

00
00

7 50
i

00

3 00 i 00
i

i2mo, Mandel's Handbook for Bio-chemical Laboratory * Martin's Laboratory Guide to Qualitative Analysis with the Blowpipe i2mo, (Considered Principally from a Sanitary Standpoint.) Mason's Water-supply. Rewritten 8vo, 3d Edition, i2n-o, Examination of Water. (Chemical and Bacteriological.) 8vo, Matthew's The Textile Fibres Meyer's Determination of Radicles in Carbon Compounds. (Tingle.). .i2mo, i2mo, Miller's Manual of Assaying i2mo, Mixter's Elementary Text-book of Chemistry. i2n:o,Morgan's Outline of Theory of Solution and its Results Elements of Physical Chemistry i2mo, lOmo, morocco, Morse's Calculations used in Cane-sugar Factories Mulliken's General Method for the Identification of Pure Organic Compounds. Vol. I Large 8vo, O'Brine's Laboratory Guide in Chemical Analysis 8vo, O'DriscoU's Notes on the Treatment of Gold Ores 8vo, (Ramsey.) Ostwald's Conversations on Chemistry. Part One. i2mo, (Turnbull.). Ostwald's Conversations on Chemistry. Part Two. (In Press.) * Penfield's Notes on Determinative Mineralogy and Record of Mircral Tests.
. .

50 60

4 00
i

25

3 50 1 00

00 50 i 00 2 co
i
i

50

5 00 2 00
^ 00
i

50

Pictet's The Alkaloids and their Chemical Constitution. pinner's Introduction to Organic Chemistry. (Austen.) Poole's Calorific Power of Fuels

(Biddle.)

Svo, paper, Svo,

50
5 00
i 50 3 00

i2mo,

8vo, Prescott and Winslow's Elements of Water Bacteriology, with Special Reference to Sanitary Water Analysis i2mo,

23

* Reisig's Guide to Piece-dyeing 8vo, 25 Richards and Woodman's Air, Water, and Food from a Sanitary Standpoint 8vo, 2 Ricbiards's Cost of Living as Modified by Sanitary Science i2mo, i Cost of Food, a Study in Dietaries i2mo, i * Richards and Williams's The Dietary Computer 8vo, i Ricketts and Russell's Skeleton Notes upon Inorganic Chemistry. (Part I. Hon-metallic Elements.) 8vo, morocco, Ricketts and Miller's Motes on Assaying 8vo, 3

00 00 00 00
50
'7S

00

Rideal's

Sewage and the Bacterial Purificat on of Sewage Disinfection and the Preservation of Food Rigg's Elementary Manual for the Chemical Laboratory
Rostoski's

8vo, 8vo, 8vo,

i^iro,

Serum Diagnosis.

(Bolduan.)

Ruddiman's IncompatibiUties in Prescriptions and Artistic Technology of Paints and VarniEh Salkowski's Physiological and Pathological Chemistry. (Orndorff.) Schimpf's Text-book of Volumetric Analysis
Sabin's Industrial

Svo, Evo,

3 50 4 00 i 2$ i 00 2 00 3 00
2 S"

Svo,
12 mo,

i2mo, Spencer's Handbook for Chemists of Beet-sugar Houses i6mo, morocco, Handbook for Sugar Manufacturers and their Chemists. i6mo, morocco,
Essentials of Volumetric Analysis
.

50 25 3 00 2 00
2
i

Stockbridge's
* Tillman's

Rocks and

Soils

Svo, Svo, Svo, Svo,

Svo,.

2
i

Elementary Lessons in Heat Descriptive General Chemistry

3 3

(Hall.) Treadwell's QuaUtative Analysis. Quantitative Analysis. (Hall.)

4
5
i

Svo, lurneaure and Russell's Public Water-suppUes r2mo, Van Deventer's Physical Chemistry for Beginners. (Boltwood.) * Walke's Lectures on Explosives Svo, Svo, Washington's Manual of the Chemical Analysis of Rocks Wassermarm's Immune Sera Haemolysins, Cytotoxins, and Precipitins. (Bol:

4
2

50 50 00 00 00 00 50 00 00
00 50 so

diian.)

i2rao,

i
t

Svo, Well's Laboratory Guide in Qualitative Chemical Analysis Short Course in inorganic Qualitative Chemical Analysis for Engineering

Students

i2mo,
Svo,

Text-book of Chemical Arithmetic. (In press.) Whipple's Microscopy of Drinking-water Wilson's Cyanide Processes
Chlorination Process Wuiling's Elementary Cotirse
in Inorganic, Pharmaceutical,

i2mo, i2mo, and Medical i2mo,

3 50 i 50 " 50
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Chemistry

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RAILWAY ENGINEERING.
i2mo, Baker's Engineers' Surveying Instruments Paper ipi X 24} inches. Bixby's Graphical Computing Table (Postage, ** Burr's Ancient and Modem Engineering and the Isthmian Canal. 8vo, 27 cents additional.) 8vo, Comstock's Field Astronomy for Engineers Svo, Davis's Elevation and Stadia Tables
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3 00

25
3 5 2 so i oo
r i

Practical

i2mo, i2mo,
Svo, Svo,
Svo,

so

00

3 00 3 S" 2 so
i 7S 3 50

i2mo,
8vo, |^' Svo,

Gore's Elements of Geodesy

Hayford's Text-book of Geodetic Astronomy Bering's Ready Reference Tables (Conversion Factors)

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2 so

Howe's Retaining Walls for Baxii^ SmaU 8vo, Johnson's (J. B.) Theory and Practice of Surveying 8vo Johnson's (L. J.) Statics by Algebraic and Graphic Methods Laplace's Philosophical Essay on Prohabilities. (Truscott and Emory.) 12010, Svo, llahan's Treatise on Civil Engineering. (1873-) (Wood.) ^^^' * Descriptive Geometry 8vo, Merriman's Elements of Precise Surveying and Geodesy
.
. .

^5 4 00 2 00 2 00 s 00
^

So

2 50
2 00 2 00

Elements of Sanitary Engineering

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for Surveyors

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Svo,
'

i2mo.
4to,

3 so 2 00
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8vo half leather.

8vo, 8vo,

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Civil Engineer's Pocket-book Wait's Engineering and Architectural Jmrisprudence

i6mo, morocco,

i6mo, morocco,
* Wheeler's Elementary Course of Wilson's Topographic Siirveying
Civil

Engineering

Svo, Svo,

BRIDGES AliD ROOFS.

Boiler's Practical Treatise

on the Construction

of Iron

Highway

Thames River Bridge

Bridges. .8vo, 4to, paper,

Burr's Course on the Stresses in Bridges and Roof Trusses, Arched Ribs, ard Suspension Bridges Svo, Burr and Falk's Influence Lines for Bridge and Roof Computations. .Svo, Du Bois's Mechanics of Engineering. Vol. II Small 4to, 4to, Foster's Treatise on Wooden Trestle Bridges Fowler's Ordinary Foundations Svo, Greene's Roof Trusses Svo, Bridge Trusses Svo, Arches in Wood, Iron, and Stone Svo, Howe's Treatise on Arches Svo, Design of Cimple Roof-trusses in Wood and Steel Svo, 2 00 Johnson, Bryan, and Turneaure's Theory and Practice in the Lcsiprirf of Modern Framed Structures Email 410, 10 00 Herriman and Jacoby's Text-book on Roofs and Bridges: Part I. Stresses in Simple Trusses Svo, 2 50 Part II. Graphic Statics Svo, 2 50 Part III. Bridge Design Svo, 2 50 Part IV. Higher Structures Svo, 2 50 Morison's Memphis Bridge 4tb, 10 00 Waddell's De Pontibus, a Pocket-book for Bridge Engineers. i6mo, morocco, 3 co Specifications for Stpel Bridges r2mo, i 25 Wood's Treatise on the Theory of the Construction of Bridges and Roofs. .Svo, 2 00 Wright's Designing of Draw-spans Part I. Plate-girder Draws Svo, 2 50 Part II. Riveted-truss and Pin-connecled Long-span Draws Svo, 2 50 " parts in one voliime. Two co
. .

".

'


Sabin's Industrial

and

Atlisliu ^ci-iiuuiUKj

t..

^iwi.o .iiu

.*i^*o..

w.o,
i2ino,

3 00
i

Smith's Materials of Machines Snow's Principal Species of Wood Spalding's Hydraulic Cement

00

8vo,

Text-book on Roads and Pavements. Taylor and Thompson's Treatise on Concrete, Plain and Reinforced

i2mo, i2mo,
8vo,

3 50 2 00
2 00 s 00

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.
.

8 00
2

00

3 50
2 50

5 00 4 00 3 00 i 25
2 00

3 00

4 00

RAILWAY ENGINEERING.
Andrew's Handbook for Street Railway Engineers Berg's Buildings and Structures of American Railroads
Brook's Handbook of Street Railroad Location
Butt's Civil Engineer's Field-book

3x5 inches, morocco,

4to,

Crandall's Transition Curve

i6mo, morocco. i6mo, morocco, i6mo, morocco.

Railway and Other Earthwork Tables 8vo, Dawson's "Engineering" and Electric Traction Pocket-book. .i6mo, morocco, Dredge's History of the Pennsylvania Railroad: C1879) Paper, * Drinker's Tunnelling, Explosive Compounds, and Rock DriUs.4to, half mor., Fisher's Table of Cubic Yards Cardboard, Godwin's Railroad Engineers' Field-book and Explorers' Guide. i6mo, mor., Howard's Transition Curve Field-book i6mo, morocco, Hudson's Tables for Calculating the Cubic Contents of Excavations and Embankments 8vo, Molitor and Beard's Manual for Resident Engineers i6mo,
. .

I i

Nagle's Field
Philbricfc's

Manual for Railroad Engineers Field Manual for Engineers

Searles's Field Engineering

Railroad Spiral Taylor's Prismoidal Formulae and Earthwork 8vo, * Trautwine's Method of Calculating the' Cube Contents of Excavations and Embankments by the Aid of Diagrams 8vo, The Field Practice of Laying Out Circular Curves for Railroads.

i6mo, morocco, i6mo, morocco i6mo, morocco, i6mo, morocco,

3
3
i

50

00 50

i2mo, morocco,
Cross-section Sheet

Paper

Webb's Railroad Construction Wellington's Economic Theory of the Location

i6mo, morocco!
of

Railways

Small 8vo,

2s 5 00 s 00

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Barr's Kinematics of Machinery * Bartlett's Mechanical Drawing " " " *

Abridged Ed

CooUdge's Manual of Drawing Coolidge and Freeman's Elements of General Drafting for Mechanical Engi-

gyp g^o' gvo, 8^o_ papg^

4to,

2 so ^ ^^
i
i

so 00

"""=
Dudley's Kinematics of Machines Emch's Introduction to ProiectivR Gpnmptrv an/i
j+e A.^i:

Oblong

2 so
,

g^p
+:

o_-'

Hill's

Text-book on Shades and Shadows, and Perspective Jamison's Elements of Mechanical Drawing Jones's Machine Design: Part I. Kinematics of Machinery Part II. Form, Strength, and Proportions of Parts MacCord's Elements of Descriptive Geometry Kinematics; or. Practical Mechanism Mechanical Drawing
Velocity Diagrams
* Mahan's Descriptive Geometry and Stone-cutting Industrial Drawing. (Thompson.)

8vo, 8vo, 8vo, 8vo,

8vo,

2 00 2 50

so

3 00 3 00 s 00 4 00 i 50 i so 3 50 s 00
2

8vo,
4to,

8vo, 8vo, 8vo,

4to,

Moyer's Descriptive Geometry. (In press.) Reed's Topographical Drawing and Sketching
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Drawing 8vo, Text-book of Mechanical Drawing and Elementary Machine Design. 8vo, Robinson's Principles of Mechanism 8vo, Schwamb and Merrill's Elements of Mechanism 8vo, Smith's Manual of Topographical Drawing. (McMillan.) 8vo, Warren's Elements of Plane and Solid Free-hand Geometrical Drawing, i2mo. Drafting Instruments and Operations i2mo. Manual of Elementary Projection Drawing i2mo, Manual of Elementary Problems in the Linear Perspective of Form and Shadow i2mo, Plane Problems in Elementary Geometry i2mo. Primary Geometry i2mo.
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00

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Small Svo, Anthony and Brackett's Text-book of Physics. (Magie.) i2mo, Anthony's Lecture-notes on the Theory of Electrical Measurements.
. . .

3
i
t

Svo, Benjamin's History of Electricity Svo, Voltaic Celt (Boltwood.).8vo, Classen's Quantitative Chemical Analysis by Electrolysis. Svo, Crehore and Squier's Polarizing Photo-chronograph Dawson's "Engineering" and Electric Traction Pocket-book. i6mo, morocco,
Dolezalek's

3 3 3

>

3 s
2
1

Theory of the Lead Accumulator (Storage Battery).

Ende.)

(Von i2mo,
Svo,

Duhem's Thermodynamics and Chemistry. (Burgess. ) Flather's Dynamometers, and the Measurement of Power
Gilbert's

4
3 2
i

i2mo,
Svo,

De Magnete.

(Mottelay.)

i2mo, Hanchett's Alternating Currents Explained i6mo, morocco, Hering's Ready Reference Tables (Conversion Factors) Svo, Holman's Precision of Measurements .Large Svo, Telescopic Mirror-scale Method, Adjustments, and Tests. .Svo, Kinzbrunner's Testing of Continuous-Current Machines Svo, Landauer's Spectrum Analysis. (Tmgle.) Le Chatelien's High-temperature Measurements. (Boudouard Burgess.) i2mo, Liib's Electrolysis and Electrosynthesis of Organic Compounds. (Lorenz.) i2mo,
. .

2 2 2
-

".

3 3
i

Wolff's Windmill as a Prime Mover 8vo, Wood's Rustless Coatings: Corrosion and Electrolysis of Iron and Steel. .8vo,

<

MATHEMATICS.
Baker's Elliptic Functions * Bass's Elements of Differential Calculus Briggs's Elements of Plane Analytic Geometry
8vo,

Compton's Manual

of Logarithmic

Computations

i2mo, i2mo, i2mo,

8vo,

i 50 4 00 i 00

Davis's Introduction to the Logic of Algebra * Dickson's College Algebra

* Introduction to the Theory of Algebraic Equations Emch's Introduction to Projective Geometry and its Applications Halsted's Elements of Geometry Elementary Synthetic Geometry Rational Geometry

Large i2mo, Large i2mo,

8vo, 8vo, 8vo,

50 50 50 i 23 2 50
i

i
1

i i i

i2mo,

7S so 7S

Vest-pocket size. paper, 100 copies for * Mounted on heavy cardboard, 8X 10 inches, 10 copies for Johnson's (W. W.) Elementary Treatise on Difierential Calculus .SmahSvo, Johnson's (W. W.) Elementary Treatise on the Integral Calculus. Small 8vo, Johnson's (W. W.) Curve Tracing in Cartesian Co-ordinates i2mo, Johnson's (W. W.) Treatise on Ordinary and Partial Differential Equations. Small 8vo, Johnson's (W. W.) Theory of Errors and the Method of Lea^t Squares. i2mo, * Johnson's (W. W.) Theoretical Mechanics i2mo, Laplace's Philosophical Essay on Probabilities. (Truscott and Emory.) i2mo, * Ludlow and Bass. Elements of Trigonometry and Logarithmic and Other Tables 8vo, Trigonometry and Tables published separately Each, * Ludlow's Logarithmic and Trigonometric Tables 8vo, Maurer's Technical Mechanics ^....8*,, Merriman and Woodward's Higher Mathematics 8vo, Merriman's Method of Least Squares 8vo, Rice and Johnson's Elementary Treatise on the Differential Calculus. Sm. 8vo, Differential and Integral Calculus. 2 vols, in one Small 8vo, Wood's Elements of Co-ordinate Geometry 8vo, Trigonometry: Analytical, Plane, and Spherical i2mo,
. . .

* Johnson's (J, B.) Three-place Logarithmic Tables:

is 5 00

2s
2 00 3 00
1 i

50 00

3 So i so 3 00
2 00 3 00 2 00
i 00 4 00 5 00 2 00 3 00 2 50 2 00 i 00

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MATERIALS OF ENGINEERING, STEAM-ENGINES AND BOILERS.
Bacon's Forge Practice Baldwin's Steam Heating for Buildings
Barr's Kinematics of Machinery * Bartlett's Mechanical Drawing

i2mo, i2mo,
8vo, Svo, Svo,

50

2 50 2 50

3 00
2

Abridged Ed Benjamin's Wrinkles and Recipes Carpenter's Experimental Engineering Heating and Ventilating Buildings Gary's Smoke Suppression in Plants using Bituminous Coal.
*
'.*

"

"

i2mo,
Svo,
Svo,
(In Prepara-

50 00 6 00 4 00
i

tion.)
Clerk's

Gas and Oil Engine

Small Svo,
Svo, paper,
of General Drafting for

4
i

Coolidge's

Coolidge

Manual of Drawing and Freeman's Elements

00 00 50

gineers

Mechanical EnOblong 4to,

11

"
o,

Cromwell's Treatise on T Treatise on Belts and Pulleys Durley's Kinematics of Machines Flather's Dynamometers and the Measurement

5a
50-

i2mo,
Svo,
of

Power

Rope Driving Gill's Gas and Fuel Analysis

Hall's

for Engineers

Car Lubrication

Bering's Ready Reference Tables (Conversion Factors) Huttou's The Gas Engine Jamison's Mechanical Drawing Jones's Machine Design Part I, Kinematics of Machinery Part n. Form, Strength, and Proportions of Parts

i2mo, i2mo, i2mo, i2mo, i6mo, morocco,

8vo, 8vo,
.8vo,

4 00 3 oa 2 00
i
i

25 00

50
50

5 00
2

50

8vo,

3 00

i6mo, morocco, 5 00 Kent's Mechanical Engineers' Pocket-book 8vo, 2 00 Kerr's Power and Power Transmission (In press.) I^onard's Machine Shop, Tools, and Methods. Lorenz's Modern Refrigerating Machinery. (Pope, Haven, and Dean.) (In press.) MacCord's Kinematics; or. Practical Mechanism 8vo, 5 00 Mechanical Drawing 4*0, 4 00
Velocity Diagrams Mahan's Industrial Drawing. 8vo,
i

50

(Thompson.)

8vo, 8vo, 8vo,

Poole's Calorific Power of Fuels Reid's Course in Mechanical Drawing Text-book of Mechanical Drawing

3 50 3 00
2

00
50

and Elementary Machine Design. 8vo,

i2mo,
8vo,
8vo,

3 00
i

Richard's Compressed Air Robinson's Principles of Mechanism

00

Schwamb and

Merrill's

Elements of Mechanism

3 00 3 00 3 00 i 00
7 503 3
2

Smith's Press-working of Metals Thurston's Treatise on Friction and Lost

Work

8vo, in Machinery and Mill

8vo,
.

Work

Animal as a Machine and Prime Motor, and the Laws of Energetics 1 2mo, Warren's Elements of Machine Construction and Drawing 8vo, Weisbach's Kinematics and the Power of Transmission. (Herrmann
Klein.)

Machinery of Transmission and Governors. Wolff's Windmill as a Prime Mover

(Herrmann

Klein.)- 8vo,
8vo,

8vo,

00
00
50

5 00

Wood's Turbines

8vo

MATERIALS OF ENGINEERING.
Bovey's Strength of Materials and Theory of Structures Burr's Elasticity and Resistance of the Materials of Engineering.
Reset.
:

8vo,

50

6th Edition.
8vo,
8vo,

Church's Mechanics of Engineering Johnson's Materials of Construction Keep's Cast Iron Lanza's AppUed Mechanics Martens's Handbook on Testing Materials. (Henning.) Merriman's Text-book on the Mechanics of Materials Strength of Materials
Metcalf's Steel.

8vo, 8vo, 8vo, 8vo, 8vo,

7 50 6 00 6 00
2

50

7 50 7 50 4 00
'

A manual for

Steel-users

i2mo, i2mo.

OO'

2 00

and Artistic Technology of Paints and Varnish 8vo, Smith's Materials of Machines i2mo, Thurston's Materials of Engineering 3 vols., 8vo, Part 11. Iron and Steel 8vo, Part in. A Treatise on Brasses, Bronzes, and Other Alloys and their Constituents 8vo, Text-book of the Materials of Construction 8vo,
Sabin's Industrial

3 00 i 00 8 00 3 50
2 50

5 00

food's (De

V.) Treatise on the Resistance of Materials the Preservation of Timber

a*i.

-n Appendix on
8vo, 8vo,
2

00

Wood's (De V.) Elements of Analytical Mechanics Wood's (M. P.) Rustless Coatings: Corrosion and
Steel

3 00

Electrolysis of Iron

and
Svo;

4 00

STEAM-ENGINES AND BOILERS.

Berry's Temperature-entropy Diagram i2mo, i2mo, Carnot's Reflections on the Motive Power of Heat. (Thurston.) Dawson's " Engineering " and Electric Traction Pocket-book .... i6mo, mor.,
i i

5
i

Ford's Boiler Making for Boiler Makers Goss's Locomotive Sparks

iSmo,
Svo,

2 2 s

Hemenway's Indicator

i 50 00 MacCord's Meyer's Modern Locomotive Construction 4to, 10 00 i2mo, i 50 Peabody's Manual of the Steam-engine Indicator Tables of the Properties of Saturated Steam and Other Vapors Svo, i 00 Thermodynamics of the Steam-engine and Other Heat-engines Svo, s 00 Valve-gears for Steam-engines Svo, 2 50 Peabody and Miller's Steam-boilers Svo, 4 00 -Pray's Twenty Years with the Indicator Large Svo, 2 50 Pupin's Thermodynamics of Reversible Cycles in Gases and Saturated Vapors. (Osterberg.). i2mo, i 25 Reagan's Locomotives: Simple Compound, and Electric i2mo, ^ 50 Rontgen's Principles of Thermodynamics. (Du Bois.) Svo, s 00 Sinclair's Locomotive Engine Running and Management i2mo, 2 00 Smart's Handbook of Engineering Laboratory Practice i2mo, 2 50 Snow's Steam-boiler Practice Svo, 3 00

Practice and Steam-engine Hutton's Mechanical Engineering of Power Plants Heat and Heat-engines Kent's Steam boiler Economy. Kneass's Practice and Theory of the Injector
Slide-valves

Economy

i2mo,
Svo,

Svo, Svo, Svo, Svo,

5
4 2

25 50 00 00 00 00 00 00 00

Spangler's Valve-gears

Svo,

2
i

Notes on Thermodynamics Spangler, Greene, and Marshall's Elements of Steam-engineering Thurston's Handy Tables

i2mo,

Svo, 3 Svo, i Manual of the Steam-engine 2 vols., Svo, 10 Part I. History, Structure, and Theory Svo, 6 Part H. Design, Construction, and Operation Svo, 6 Handbook of Engine and Boiler Trials, and the Use of the Indicator and the Prony Brake Svo, 3 00 Stationary Steam-engines Svo, 2 50 Steam-boiler Explosions in Theory and in Practice i2mo, i 50

50 00 00 50 00 00 00

Manual

of Steam-boilers, their Designs, Construction, and Operation Svo, Weisbach's Heat, Steam, and Steam-engines. (Du Bois.) Svo, Whitham's Steam-engine Design Svo, Wilson's Treatise on Steam-boilers. (Flather.) i6mo, Wood's Thermodynamics, Heat Motors, and Refrigerating Machines. .Svo,
.

5 00 s 00 s 00 2 50

4 00

MECHANICS AND MACHINERY.

Barr's

Kinematics

of

Machinery

Svo, Svo,

2 50
7 50

Bovey's Strength of Materials and Theory of Structures Chase's The Art of Pattern-making Church's Mechanics of Engineering

i2mo,
Svo,

50 6 00
2

13

i2mo, :2mo, i2mo, Treatise on Belts and Pulleys Dana's Text-book of Elementary Mechanics for Colleges and Schools. .i2mo, i2mo, Dingey's Machinery Pattern Making Dredge's Record of the Transportation Exhibits Euilding of the World's .4to half mcrocco, Columbian Exposition of 1803 Du Bois's Elementary Principles of Mechanics 8vo, Vol. L Kirematics
Vol.
II.

Church*s Notes and Exai^Ti^it^s i^: ^^.--^li^r...-^Compton's First Lessons in Metal-working. Compton and De Groodt's The Speed Lathe Cromwell's Treatise on Toothed Gearing

vo,
i2n:o,

2
i
i *

00
50 50 50

i
i

50

s&
00

5 oo^
3 50 4 00 3 50 7 50 10 00 4 00 i 00

Statics

Vol. III.

Kinetics.

8vo, 8vo,
Vol.
I

Mechanics

of Engineering.

Vol. II

Small 4to, Small 4to,

8vo,
of

Durley's Kinematics of Machines

Fitzgerald's Boston Machinist

Dynamometers, and the Measurement Rope Driving Goss's Locomotive Sparks

Flather's

Power

i6mo, i2mo, i2mo,

8vo,

3 00
2 oo^
2
i

Hall's Car Lubrication

Holly's Art of

Saw

Filing

i2mo, i8mo,

00 00
75

James's Kinematics of a Point and the Rational Mechanics of a Particle. (In press.) * Johnson's (W. W.) Theoretical Mechanics i2mo, 3 00 Johnson's (L, J.) Statics by Graphic and Algebraic Methods 8vo, 2 00 * Machine Design Jones's Part I. Kinematics of Machinery 8vo, i 50 Part II. Form, Strength, and Proportions of Parts 8vo, 3 00 Kerr's Power and Power Transmission 8vo, 2 00 Lanza's Applied Mechanics 8vo, 7 50' Leonard's Machine Shop, Tools, and Methods. (In press.) Lorenz's Modern Refrigerating Machinery. (Pope, Haven, and Dean.) (In press.) MacCord's Kinematics; or. Practical Mechanism 8vo, 5 00
Velocity

Diagrams

8vo,

Maurer's Technical Mechanics 8vo, Merriman's Text-book on the Mechanics of Materials 8vo, * Elements of Mechanics i2mo, * Michie's Elements of Analytical Mechanics 8vo, Reagan's Locomotives: Simple, Compound, and Electric i2mo, Reid's Course in Mechanical Drawing 8vo, Text-book of Mechanical Drawing and Elementary Machine Design. 8 vo, Richards's Compressed Air i2mo, Robinson's Principles of Mechanism 8vo, Ryan, Norrls, and Hoxie's Electrical Machinery. Vol. 1 8vo, Schwamb and Merrill's Elements of Mechanism 8vo, Sinclair's Locomotive-engine Running and Management i2mo, Smith's (O.) Press-working of Metals 8vo, Smith's (A. W.) Materials of Machines i2mo, Spangler, Greene, and Marshall's Elements of Steam-engineering 8vo, Thurston's Treatise on Friction and Lost ^7ork in Machinery and Mill

i 50 4 00 4 00 i 00

4 00 2 50 2 00
3 00
i
^30

3 00
2 50 3 00

2 00 3 00 i 00 3 00

Work

Animal

as

ti

Machine and Prime Motor, and tte Laws

8vo, of Energe'ics.

3 00
X

i2mo,
Warren's Elements of Machine Constructior. and Drawing Weisbach's Kinematics and Power of Transmission. (Herrmann Machinery of Transmission and Governors. (Herrmann Wood's Elements of Analytical Mechanics Principles of Elementary Mechanics
Turbines

00

Klein. 8vo, Klein.). 8vo,

)
.

8vo,

7 50 S 00 5 00 3 00
i

8vo,

i2mo,
gvo.
4to,

2
1

The World's Columbian Exposition

of 1893

25 50 00

METALLURGY.
Egleston's Metallurgy of Silver, Gold, Vol. I. Silver
Vol. II.

and Mercury:
8vo, 8vo,
i2nio.
7

Gold and Mercury

** Iles's Lead-smelting. (Postage 9 cents additional.) Keep's Cast Iron Kunhardt's Practice of Ore.Dressing in Europe

Le

Chatelier's High-temperature Measuremepts.

(Boudouard

Burgess. )i2nio,

8vo, Svo,

Metcalf's Steel.

Manual

for Steel-users-

izmo,

Smith's Materials of Machines i2mo, Thurston's Materials of Engineering. In Three Parts Svo, Part II. Iron and Steel Svo, Part ni. A Treatise on Brasses, Bronzes, and Other Alloys and their Constituents Svo, Svo, Ulke's Modem Electrolytic Copper Refining

MINERALOGY.
Barringer's Description of Minerals of Commercial Value. Boyd's Resources of Southwest Virginia

Oblong, morocco,
Svo,

Map
Brush's

of Southwest Virignia of Determinative Mineralogy.

(Penfii-^ld.)

Pocket-book form.
Svo, Svo, paper. Cloth,

Manual

Chester's Catalogue of Minerals

Dictionary of the Names of Minerals Dana's System of Mineralogy First Appendix to Dana's New " System Text-book of Mineralogy Minerals and How to Study Them

Svo,
of

Large Svo, half leather. Mineralogy." Large Svo,

Svo,

i2mo.

Large Svo, i2mo Mineralogy and Petrography i2mo, Douglas's Untechnical Addresses on Technical Subjects Svo, Eakle's Mineral Tables Svo, Eglpston's Catalogue of Minerals and Synonyms Hussak's The Determination of Rock-forming Minerals. (Smith.). Small Svo, Merrill's Non-metallic Minerals; Their Occurrence and Uses Svo, * Penfield's Notes on Determinative Mineralogy and Record of Mineral Tests, Svo paper, Rosenbusch's Microscopical Physiography of the Rock-making Minerals.

Catalogue of American Localities of Minerals

Manual

of

Important Minerals and Rocks Williams's Manual of Lithology

(Iddings.) * Tillman's Text-book of

Svo. Svo. Svo,

MINING.
Beard's Ventilation of Mines i2mo, Boyd's Resources of Southwest Virginia Svo, Map of Southwest Virginia Pocket book form. Douglas's Untechnical Addresses on Technical Subjects i2mo. * Drinker's Tunneling, Explosive Compounds, and Rock Drills 4to, hf mor. Eissler's Modern High Explosives Svo Fowler's Sewage Works Analyses .i2mo Goodyear's Coal-mines of the Western Coast of the United States i2mo.
. . . .

Ihlseng's

Manual

of

Mining

Svo,

**

Lead-smelting. (Postage gc. additional.) Kunhardt's Practice of Ore Dressing in Europe O'Driscoll's Notes on the Treatment of Gold Ores * Walke's Lectures on Explosives Wilson's Cyanide Processes Chlorinatlon Process.
Iles's

i2mo,
Svo,

Svo. Svo,

i2mo, i2mo,

15

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