BIOLOGY PRACTICAL 1

FIS 1082

PRACTICAL 2: MICROSCOPY

NAME: THEVAMALAR D/O NADARAJA

ID NO: 981022-04-5420
RACTICAL DATE: 2.10.2019
SUBMIT DATE: 9.10.2019
LECTURER: MOHD SHAHMEN BIN MOHD YAZAM
2.1 OBJECTIVE:
1. Identify each of microscope components and their function.
2. Practice the operating procedure of light microscope
3. Perform wet mount procedure.

2.2 INTRODUCTION:
Light microscope is one of the most important equipment that is used in microbiology
laboratory. Correct usage of the microscope will speed up the examination process and
also determine the level of successful of an equipment. Below are the proper ways to use
a microscope.
1. Always ensure the slide is clean and dry before placing it on the stage. Ensure the
lenses are free from dust.
2. After using the oil immersion lens, ensure the lens is free from oil reside before
you store it. Always use lens cleaning paper to clean the lens.
3. When handling a microscope, hold the limb with right hand and use the other hand
to support from the bottom. It must be carried vertically whenever possible.
A) MICROSCOPE COMPONENT
2.3 MATERIALS / APPARATUS:
1. Light microscope

2.4: METHODOLOGY:
The components of microscope is labelled below.

B) USE OF MICRSCOPE
2.5 MATERIAL / APPARATUS:
1. Histology slides
2. Glass slide
3. Cover slip
4. Lens cleaning paper
5. Immersion oil
6. 90% alcohol
7. Distilled water
8. Tissue roll
9. Cutter / Scissors
10. Disposable dropper
2.6 METHODOLOGY:
Wet mount procedure
1. A drop of water is placed on the center of the slide. It can be placed the specimen
(words) at first on the slide, but small specimen usually separated more easily from
the tweezers or needle it dipped into the drop of water.
2. The specimen was placed into the drop of water and another drop is added on it
when the specimen float. That was to reduce the possibilities of air bubbles
forming.
3. Cover glass was lowered carefully and it touched with one side the drop of water.
The cover slip formed an angle of 45 degrees with the slide. The cover glass was
touched on slide to prevent finger prints. Alternatively tweezers is used to hold the
cover glass.
4. Cover slip lowered completely. Cover slip was placed at an angle to prevent the
formation of air-bubbles.
5. Excess water was removed with a filter or tissue paper.

Microscope Observation Procedure

1. The revolving turret/ nosepiece was turned so that the lowest power objective lens
(eg.4x) can be clicked into position.
2. The microscope slide was placed on the stage and fasten it with the stage clips
3. Looked at the objective lens and the stage from the slide and focus knob was
turned so the stage moves upward. Is was moved up as far as it will go without
letting the objective touch the coverslip.
4. The image was looked through the eyepiece/ocular lens and move the focus knob
until image comes into focus.
5. The condenser and light intensity was adjusted for the greatest amount of light.
6. The microscope slide was moved around until the sample was in the center of the
field of view. (what you see)
7. The focus knob was used to place the sample into focus and the condenser is
readjusted and also the light intensity for the clearest image (with low power
objective you might need to reduce the light intensity or shut the condenser).
8. Clear image of our sample has been shown with lowest power objective. The
objective lens is changed to next magnification. We might needed to readjust the
sample into focus and readjust the condenser and light intensity. Steps 3 through
5 is repeated when there the image has not focused on specimen with high power
objective lens in place. The objective lens was not touched the slide.
9. Once finished, the stage is lowered, the lower power lens is clicked into position
and the slide has been removed.
 2.7 RESULTS:

A. Eyepiece
B. Nose piece
C. Objective lens
D. Stage
E. Illumination
F. Base
G. Arm
H. Coarse adjustment
I. Fine adjustment
J. Stage control
K. Light intensity control
2.8 DISCUSSION
Microscope is the object which help to see small things or micro things that cannot be
seen with our naked eyes. There are 6 types of microscopes with different functions
respectively. Those microscope have different objective lenses that magnify the image
with the refraction of light intensity. Always start with the lower power of light intensity and
lower magnification. Then adjust the knob gently until get clearer image. Use the glass
slide carefully as it is a sharp object. Use immersion oil while using high magnification of
100x100 to get a clearer image. Place the cover slip at angle of 45 degrees to prevent
forming of water bubbles.

2.9 CONCLUSION:
1. The components of microscope and its functions are learned.
2. Practical how to operate of light microscope.
3. Wet mount procedure has performed.

2.10 QUESTIONS:
1. List down types of microscope.
a) Fluorescence Microscope
b) Electron microscope; Transmission Electron Microscope (TEM) and Scanning Electron
Microscope (SEM)
c) Phase Contrast Microscope
d) Surgery Microscope
e) Light Compound Microscope
f) Acoustic Microscope, Stereomicroscope Confocal Laser Scanning Microscope, Total
Internal Reflection Fluorescence Microscope (TIRF).
2. List the component of microscope and their function.
COMPONENT FUNCTION
A. Eyepiece The lens where the viewer look for the
specimen under microscope
B. Nose piece A rotating turret that adjust the objective
lenses.
C. Objective lens Which have different magnification of
lenses (4x10, 10x10, 40x10, and
100x100) to look the specimen.
D. Stage The platform where slide is placed.

E. Illumination Light source for the microscope.

F. Base Where the illumination is located and it

give support for the microscope.
G. Arm The frame where the base and body
connected in microscope,
H. Coarse adjustment Focus the specimen generally.

I. Fine adjustment Focus the specimen in detail.

J. Stage control Control the position of the slide.

K. Light intensity control Provide light to the specimen.

3. Why do we need to use immersion oil in order to observe (using 100x lens)?
The light refraction is high when use high magnification lens (100x). So immersion
oil needed to give a clearer image.
4.Do you experience any problem while observing the image of the specimen? Explain
briefly the solution you did.
While observing image at 40x magnification it took time to get a clearer image. To
get a better image use fine focus control on a slide by adjusting the knob gently.
2.11 REFERENCES:
1. https://www.microscopemaster.com/parts-of-a-compound-microscope.html
2. Lab manual Biology Practical 1