Microscopy

Experiment 1
MICROSCOPY
OBJECTIVES

At the end of this experiment, the students should be able to:

1. identify the parts and uses of a compound microscope
2. learn to adjust the microscopes for viewing biological specimens
3. learn to measure small objects using the principles of microscopy

MATERIALS
To be assigned per group: From the preparation room:
newspaper compound microscope
pencil microscope slides & Cover slips
scissors prepared slides (Euglena, Allium cepa, Paramecium)
ruler
medicine dropper

INTRODUCTION

The microscope is the biologist’s basic tool. It was developed to explore living things that are too small
to be seen with the naked eye.
Early microscopes, like the one Leeuwenhoek made, had only one lens and were difficult to use. The
biggest problem with this kind of microscope was magnification. The more powerful the lens for magnification
the closer the viewer’s eye had to be to the lens. A major advancement in microscopes came with the invention
of the compound microscope. This type of microscope has two sets of lenses, which magnify objects much
more than a single lens.
The compound microscope has four basic parts: the lens system, the focusing system, and the lighting
system. The lens system has two sets of lenses that are called objective lenses. These lenses work as much as
did the lens of the early, simple microscope. The objective lenses make the initial or primary magnification.
They are located in the nosepiece of the microscope. Inscribed on each objective is the magnification or power
of that lens. This tells the number of times the lens magnifies the image. The second kind of lens in the
microscope is the ocular lens, which is sometimes called the eyepiece. This is located at the top of the body
tube. The ocular serves as a small telescope and magnifies the image made by the objective lens. This
enlargement is called as the secondary magnification.
A laboratory specimen to be viewed through the microscope is mounted on a glass slide and covered
with a cover slip. The slide rests on a stage, the flat surface beneath the body tube. Stage clips hold the slide in
place. Also, they help in making slight adjustments in the slide’s position by holding the slide steady.
For the viewer to see the specimen, light must pass through it and the lenses to your eye. The lighting
system is located under the stage of the microscope. There are three types of lighting system. The simplest
system uses a concave mirror that focuses a beam of light on the slide. The second type of lighting system uses
a lens under the stage to focus the light. The flat side of the mirror which is found under the stage is used to
reflect light through this lens .The third type of lighting system uses a sub stage light instead of a mirror. Its
disadvantage is that light gets hot and can easily destroy the specimen.
The focusing system brings the image of the specimen into proper focus. It is necessary to change the
distance between the slide and the objective lens to accomplish proper focusing. This can be done in two ways,
depending upon the microscope you are using. Either the lens can be moved or the stage upon which the slide
rests can be moved .There are also two knobs that are used to control the focus. These are the coarse and fine
adjustment knobs.
In this experiment, the structure, adjustments and measurements in microscopy will be studied. The
students will have hands on experience in the use of compound microscopes in the laboratory.

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Microscopy

PROCEDURE

A. Parts and Functions of the Compound Microscope

1. Obtain a compound microscope and prepare a labeled diagram in your laboratory report sheet.
2. Identify the parts and uses of your labeled microscope.

B. Magnification and Power of the Compound Microscope

1. Put the low power objective in place. Look through the ocular and adjust the light so that you see a
uniformly bright field of view.
2. Prepare a slide to be viewed under the microscope. Cut a lowercase “e” from a newspaper and place it
in the center of a clean side. Put a drop of water under the letter.
3. Place the edge of a cover slip against the water and with a pencil gently lowers the cover slip over the
“e”. (Placing the cover slip in this manner prevents bubbles from forming. This is also called wet
mount.)
4. Place the slide under the stage clips, so that the “e” is right side up. Focus the “e”.
5. Focus the “e” with the low power objective (10X) first by clicking the low power objective into position
in the nosepiece.
6. Turn the coarse adjustment knob until the objective is as close as possible to the slide without touching
it.
7. Look through the ocular and turn the coarse adjustment knob in the direction that will move the object
away from the stage. The “e” will come into approximate focus. To sharpen the focus, turn the fine
adjustment knob back and forth.
8. Look at “e’ using the high power objective by switching the nosepiece. Sharpen the focus by turning the
fine adjustment knob. (If you cannot find “e” under high power, look through the ocular and move the
slide slightly or move the slide in other directions.)
9. When you are finished using the microscope, remove the slide from the stage. Rinse the cover slip and
the slide with water. Dry the slides.
10. Return the microscopes and remember to carry them with one hand under the base and the other
holding the arm.

C. Measurement Using the Compound Microscope

1. Using the 10X objective in place put a ruler across the middle of the stage and focus on the millimeter
markings. The unit usually used in microscopic measurement under the compound microscope is the
micrometer (µm). The micrometer is one-thousandth of a millimeter.
2. Rotate the nosepiece to bring the high power objective into position.
3. Calculate the diameter of the field under high power by dividing the magnification power of the HPO by
the magnification of the LPO. The diameter of the low power field can be obtained by dividing the value
of the LPO with the value of the high power field. This value is the approximate diameter of the high
power field.

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NAME (SN, GN, MI) ______________________________ DATE PERFORMED ____________________________

SUBJECT/SECTION ______________________________ DATE SUBMITTED ____________________________
INSTRUCTOR ___________________________________ RATING ____________________________________

Report for Experiment 1

MICROSCOPY

A. Label the parts of a compound microscope below.

B. Questions on Magnification and Power of the Microscope

1. What is the magnification power of the ocular lens and each objective lens of the compound microscope
you are using?

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2. Compare the length of each lens. Which lens is the longest and why should this be?

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3. Calculate the magnification for each lens combination on the microscope. Show your calculations.
(Note: The total magnification is the product of the magnification of the ocular and the objective lens ).

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4. Describe the position of the letter viewed through the microscope and compare its position on the
stage?

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5. Compare the direction of movement of the specimen on the stage with the movement of thee image as
seen through the microscope.

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Questions on Measurements in Microscopy

1. What is the diameter of the field of your compound microscope in micrometer with the 10X objective in
place? (Show your calculation including conversion factors )
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2. Calculate the diameter of the high power field of view on your microscope.
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C. Conclusions
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D. Applications

1. What are the two kinds of lenses on a compound microscope? What does each do?
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2. How do you control the amount of light that passes through the specimen?
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3. What is the relationship between the movement of the specimen on the stage and the movement of
image seen through the lens?
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4. How does one determine the magnification of a microscope?

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REFERENCES (In standard bibliographic format)

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