ACTIVITY NO #: 1

ACTIVITY TITLE: Microscopy

NAME: ARAO, Valerie Frances P. January 27, 2024
Objectives
• Identify the different parts of the compound microscope.
• Describe the function of each part.
• Demonstrate the correct use and care of a compound microscope.
• Be able to troubleshoot microscope.

Introduction:
Light, or optical, microscopes are
essential for histological studies because
they allow us to visualize cells and
morphological features of tissues. Light
microscopes rely on glass lenses and visible
light to magnify tissue samples. It was
invented in XVII century, and has been
improved over the years, resulting in the
powerful modern light microscopes. The most
remarkable improvement has been getting
better glass lenses to obtain sharper and
non-distorted images, as well as adding
devices to explore new ways of visualizing
tissular features.
Light microscopes contain two main
lenses: objectives and eyepieces (ocular).
Objective gathers the light that went through the tissue, whereas the eyepieces project the tissue image on
the eye. The total magnification is the result of multiplying the objective magnification by the eyepiece
magnification. For example, if we have a 40x objective (magnifies 40 times) and a 1x eyepiece (magnifies
10 times), the total magnification is 400 times. More advanced microscopes can get 1000 to 1500
magnifications (100x objective and 10x or 15x eyepiece). Some light microscopes may contain additional
internal lenses between the objective and the eyepiece that can change the total magnification.
Magnification and resolution power must not be confused, because not matter how we can magnify an
image, including digital methods, the resolution cannot be increased.
 Let’s find out how much you already know about this module. Briefly answer the following
questions.

Prelab Questions:
1. What are the three components of a microscope?

Basic Structural Components:

1. Mechanical Parts
2. Magnifying Parts
3. Illuminating Parts/System

2. How electron microscope is different from light microscope?

Light microscopes are good for observing larger structures and live specimens and is
mostly used when miniscule details aren't needed, while electron microscopes are
essential for studying and viewing the fine details of tiny objects and the inner workings of
cells and molecules.

3. Why is that microscopy is essential to histopathology?

Because microscopy is the eyes of Histopathology. Microscopes give a different and

detailed view of tissues by magnifying cells and their intricate arrangements. With that,
pathologists can spot abnormalities which is essential in the medicine field and to the
overall health stability of a community. Without microscopy, histopathology will not
progress, and diagnosing and treating diseases would be a much greater challenge.

Materials:
Compound Microscope
Prepared tissue sample (Colon or Gall Bladder)

Procedure
1. Place the microscope on the bench squarely in front of you.
2. Obtain a prepare slide and place it in the clips on the mechanical stage.
3. Adjust the eyepieces on a binocular microscope to your own personal measurements.
a. Look through the eyepieces and using the thumb wheel, adjust the distance between the
eyepieces until one circle of light appears.
b. With the low-power (10x) objective in place, cover the left eyepiece with a small card and focus
the microscope on the slide. When the right eyepiece has been focused, remove your hand
from the focusing knobs and cover the right eyepiece. Looking through the microscope with
your left eye, focus the left eyepiece by turning the eyepiece adjustment. Make a note of the
number at which you focused the left eyepiece so you can adjust any binocular microscope for
your eyes.
4. Raise the condenser up to the stage. On some microscopes, you can focus the condenser by the
following procedure:
a. Focus with the 10X objective.
b. Close the iris diaphragm so only a minimum of light enters the objective lens.
 c. Lower the condenser until you see the light as a circle in the center of the field. On some
microscope, you can center the circle of light (Figure 4) by using the centering screws found on
the condenser.
d. Raise the condenser up to the slide, lower it, and stop when the color on the periphery changes
from pink to blue (usually 1 or 2 mm below the stage).
5. Open the iris diaphragm until the light just fills the field.
6. Adjust the contrast by changing the diaphragm opening.
7. When you brought an image into focus with high power, rotate the turret to the next lens, and the
subject will remain almost in focus. All of the objectives (with the possible exception of the 4X) are
parfocal; that is, when a subject is in focus with one lens, it will be in focus with all the lenses. When
you have completed your observations under low power, swing the high-dry objective into position
and focus. Use the fine adjustment. Only a slight adjustment should be required.
8. Move the high-dry lens out of position, and place a drop of immersion oil on the area of the slide
you are observing. Carefully click the oil immersion lens into position. It should now be immersed
in the oil (Figure 5). Careful use of fine adjustment knob should bring the object into focus.
9. Record your observations and note the magnifications.

(a) (b) (c)

FIGURE 4 Focusing the condenser. (a) Using low power, lower the condenser until a distinct
circle of light is visible. (b) Center the circle of light using the centering screws. (c) Open the iris
diaphragm until the light just fills the field. Precision Graphics, Johnson, et al (2014)
 ocular lens head
Data and Results

1. Label the parts of the compound microscope .

arm

diopter adjustment
nose piece
aperture
objective lenses
stage clip stage controls

mechanical stage
course
condenser adjustment knob
illumination
fine adjustment
knob
light switch

brightness
base adjustment

2. From the labelled parts of microscope in number 1, identify which parts are under mechanical,
magnifying and illuminating parts and give its general functions.
Mechanical Parts Magnifying Parts Illuminating Parts

Base
Eyepiece/ ocular lens Condenser Lens
Light Switch
Head/Body Tube Illuminator
Course Adjustment

Fine Adjustment Turret/ Nosepiece

Brightness adjustment Objective lenses

(10x, 15x, etc.)
Arm
Aperture
Diopter adjustment

Stage controls
Stage clip

Mechanical stage
Arm: Provides support and
connects eyepiece tube to
Functions Functions Functions
Stage: Holds and positions the specimen slide.
the base. Head/Body Tube: Connects the eyepiece to the Condenser Lens: Focuses light from
Base: Provides stability and houses the illuminator. objective lenses and houses prisms for further the illuminator onto the specimen for
Aperture: Part of the magnification. proper brightness.
mechanical stage where Coarse and Fine Focus Knobs: Adjust the focus of
light can come through. the microscope. Nosepiece (Turret): Holds multiple objective Illuminator: Provides the light source
lenses with different magnifications and allows
Diopter adjustment: Adjust Light switch: Turns the illumination light on or off. switching between them. for observing the specimen.
the focus of each ocular
lenses. Brightness adjustment: Adjusts the illumination's Objective Lenses: Provide the initial
intensity. magnification of the specimen.
Stage controls: Enables the
stage to be moved back and Ocular lens: This magnifies the image of the
forth. specimen slide (generated by the objectives) for
viewing.
Stage clip: Locks and makes
sure that the specimen slides
are in place on the stage.

3. Examine the slide in the stained H&E

The drawing is in the last page due to insufficient space.

CONCLUSION

• Scanning objective: Provides a broad overview of the tissue structure, showing

the general arrangement of cells but lacking detail.
• Low power field: Offers sufficient magnification to distinguish individual cells,
revealing basic cellular shapes and boundaries.
• High power field: Functions like a zoom lens, further magnifying the cells to
showcase finer details like internal structures and surface features. It uncovers
details invisible at lower magnifications.

In conclusion the scanning objective is useful for initial orientation and assessing
gross tissue features. The low power field is deal for identifying different cell types
based on their morphology and organization within the tissue And lastly, the high
power field enables detailed examination of individual cells, including internal
structures, surface features, and potential abnormalities.

Critical Thinking and Application

1. Lisa was assigned to view the tissue sample of the small intestine under the compound
microscope. She started to operate and observe the specimen; however, she said everything
was dark, and she could not see anything. What could be the problem?
The light source/ illuminator wasn't switched on or maybe the
illuminator of the microscope she's using is faulty.

2. Your teacher asks you to help Antonio because he has a problem operating the microscope,
especially high-power objectives. He said that everything disappears every time he views the
specimen under HPO. How will you resolve Antonio's issue?
I will first check if his specimen slide is aligned properly and then teach him how to
navigate by using the fine adjustment knob. If this doesn't work, I'll check if there is a
problem with the light intensity, or if his objective lenses is dusty and in-need of
cleaning.
 3. Mirabel complained that the prepared slide given to her had no specimen to observe; she
added that it was all blurry. However, when the prepared slide was examined under a different
microscope, there was a specimen, and it was crystal clear. What could be the problem with
Mirabel's microscope?
It could be a faulty light/ illuminator or a focusing issue due to lens damage or faulty focus
knobs. Also, Mirabel might've used the HPO without navigating her area of interest using the
LPO.

REFERENCES:
Lecture notes in medical technology. http://mt-lectures.blogspot.com/2017/08/lecture-13-morphological-
examination-of.html
Atlas of plant and animal histology. https://mmegias.webs.uvigo.es/02-english/6-tecnicas/6-
optico.php#:~:text=Light%2C%20or%20optical%2C%20microscopes,light%20to%20magnify%20tissue
%20samples.
ThomasTKtungnung. (February 13, 2020). Blood smear preparation and staining practical lab.
https://www.youtube.com/watch?v=KSs0SMfERuA
Spitalnik, P.F. Histology laboratory manual. Vagelos College of Physicians & Columbia Universe