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Histology and Embryology: Dr. Ajay Kr. Mishra Jilin Medical College

Histology is the study of tissues and how they form organs. Key methods include preparing tissue sections, staining them with hematoxylin and eosin, and examining them under light and electron microscopes. Other techniques reveal the chemical composition, antigen presence, and behavior of living cells and tissues.

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Jyothi Reddy
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71 views

Histology and Embryology: Dr. Ajay Kr. Mishra Jilin Medical College

Histology is the study of tissues and how they form organs. Key methods include preparing tissue sections, staining them with hematoxylin and eosin, and examining them under light and electron microscopes. Other techniques reveal the chemical composition, antigen presence, and behavior of living cells and tissues.

Uploaded by

Jyothi Reddy
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
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Histology and Embryology

Dr. Ajay Kr. Mishra


Jilin Medical college
Introduction
Histology is a science which studies the nor
mal microstructure of the human body and th
e relationship between the body’s structure a
nd functions.

Cells are the smallest structural and functi


onal unit of human body.
Key points of this class

 Methods employed in Histology


1. Histology
 Histo=Tissue
 Logy=Study

2. Embryology

 Embryo = unborn or unhatched offspring


 Logy = study
What we will learn in this course

 Tissue: 4 basic tissues


 Organ system
Why it is important to learn histology?

 To recognize normal tissue and cells.

 Histology is the meeting-place of anatomy,


biochemistry and physiology.

 To acquire basic skill which you will use thr


oughout your career!!!
What you need to do

 Look
 Think
 Compare
 Remember
 Idealized images
How to study histology?

 There are various histological techniques most


common being Observation of histological slid
es under microscopes.

 The routine histological preparation for LM ex


amination is a paraffin section stained with H
&E.
Basic methods of histology
Procedures for making H&E preparation ar
e as follows:

Procedure:
Specimen Fixation Dehydration Clearing
Embedding Sectioning H&E staining
Observing
Obtaining the specimen

 Tissue should be fresh as possible. The tissue t


aken are cut into small pieces no more then 1x
1 cm.
Fixation solution (fixative)

Fixation
LM--- 4% formaldehyde – Fixative and
antibacterial agent

EM--- Glutaraldehyde followed by


Osmiumtetroxide
Dehydration

Fixed tissue are immersed in a series of ethyl alc


ohol solution, thereby increasing concentration
until absolute alcohol is reached.

Water in the tissue is replaced by alcohol.


Clearing
 Xylene is commonly used as a clearing agent.
Dehydrated tissue are passed through successi
ve changes of xylene until all the alcohol is rep
laced by xyeline.

 Since xyeline has a refractive index, the tissue


become transparent.
Embedding
 Tissue are embedded in the paraffin.

 Tissue are passed through two changes of melt


ed paraffin to ensure the replacement of all xyl
en with paraffin and then allowed to cool and
harden.
Sectioning
 Microtome
section thickness 3-8μm and then mounted on glass slide

Units
M, mm, μm, nm
1m= 103 mm = 106 μm= 109 nm

1μm=0.001mm=10-6m
1nm=0.001μm=10-6mm=10-9m
Microtome
Staining
 Prior to staining, paraffin section are dewaxed
and rehydrated.

 Section are then stained H&E.

 Stained section are then dehydrated through gr


aded alcohols, cleared in xylene and covered
with cover glasses.

 Frozen section and smear preprations.


Staining

 Basophilia , Acidophilia, Neutrophilia, metach


romasia
 Basic dyes: toluidine blue, methylene blue, he
matoxylin (blue)
 Acid dyes: orange G, eosin, acid fuchsin
(pink)
 common staining: Hematoxylin & Eosin
 Specific staining: silver
Hematoxylin & Eosin (H&E)

H&E is the most commonly method in histolo


gical study

Hematoxylin Blue
Eosin Pink
Hematoxylin Eosin H&E
Microscopes
 Instrument with lenses for magnifying objects or
details invisible to the naked eyes.

 Light microscope
 Electron microscope
Light microscopy

The most common type of microscope used in la


b. The device works by passing visible light th
rough a condenser and an objective lens.
Specific microscopy

(1). Phase-contrast microscope


(2). Differential interference microscope
(3). Fluorescence microscope
(4). Confocal microscope
Electron microscopy
 A microscope which beams electrons at and th
rough the object of interest instead of light bea
ms. Instead of a glass lens to bend the light, a
powerful magnet is used to bend the electron b
eam. The microscope can only be operated in a
vacuum. Provides the greatest resolution of ext
remely small details available and has been us
ed to see individual atoms.
Electron microscopy

Transmission Electron Microscopy


EM
Scanning Electron Microscopy
Transmission Electron Microscopy

 Electron beam passes through a series of electr


omagnetic lenses for focusing & magnification
instead of the glass lenses in the LM. Magnifie
d image so formed is made visible to the huma
n eye by causing the electrons to project on to
a fluorescent screen
Photograph of the transmission electron microscop
e.
Schematic view of
a transmission elec
tron microscope w
ith its lenses and th
e pathway of the el
ectrons. CCD, char
ged coupled devic
e.
Scanning Electron Microscopy
 A specimen for examination under the SEM is
coated evenly with a layer of platinum or gold.
As the electron bombards the surface of the sp
ecimen, secondary electrons are emitted from t
he surface and collected by a detector revealin
g the 3D surface architecture of cells and tissu
e
Schematic view of a scanning electron microscope.
Other methods
A. Histochemistry and cytochemistry
Reveal the chemical composition of ti
ssue and cell.

B. Immunohistochemistry and Immunocyt


ochemistry
Antigen and antibody reaction, use labeled antibodie
s as specific reagents for localizing tissue and ce
ll constituents (antigens) in situ.
Other methods (continue)
C. In situ hybridization
Is a method for detection of specific RNA or DNA sequences dire
ctly in cells or tissue section and it Use DNA or RNA probe to
detect the complementary DNA sequence.

E. Cell and tissue culture


Is a technique for the study of living cells or fragment of tiss
ue are cultured in a sterilized culture medium at appropriate te
mperature. since medium contains essential nutrients cells gro
w in vitro. Useful in detecting the effect of various reagents on
living cells.
Summary
1. Histology is the study of the tissues of the
body and of how these tissues are arranged to
constitute organs.
2. Histology=Microscopic anatomy
 3. Section preparation and H&E staining 
4. Other methods

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