ZOOLOGY LABORATORY

Exercise No. 1
THE MICROSCOPE

Background

This activity gives the students the opportunity to use a basic instrument of biology– the microscope. The
microscope enables the student to observe enlarged images of tiny objects. There are various types of
microscopes that have different functions and levels of magnification. This activity will familiarize the
students with the proper care and use of the brightfield compound microscope and dissecting microscope.

Learning objectives:
1. Identify parts of the microscope.
2. Become familiar with the use of microscopes.
3. Appreciate the importance of microscopes by observing wet mount specimen.
4. Perform basic techniques in microscopy.

Materials:

• Compound microscope
• Dissecting microscope
• Glass slides
• Cover slips
• Dropper
• Newspaper clippings
• Scissors
• One-peso coin

Procedure:

A. The Microscope and its Parts

Draw and label the parts of the microscope.

a. The eyepiece or ocular is the lens through which the object or the specimen is viewed. This lens magnifies
the image passing through it by 10 times. It also has a pointer which may be used to indicate the object or
part of an object being viewed.

b. The revolving nosepiece holds four objective lenses. Grasp the nosepiece and note that it may be rotated,
bringing each objective lens into place. Feel the distinctive "click" as each objective is in proper position
for viewing.

c. The objective lens gives the initial magnification. In general, the lower the magnification, the shorter is
the objective lens and the farther is it positioned from the specimen when focu
sed. The microscope may have four of these, located on the revolving nosepiece. These are as follows:

• The scanning lens is a short lens which magnifies the size of an object 4 times. It has the broadest
field of view of all the lenses and is used for initial focusing of objects.
• The low-power objective (LPO) magnifies by 10 times.
• The high-power objective (HPO) magnifies by 40 times. It is much longer than the first two.
 • The longest objective is the one with 100x magnification. This is the oil immersion objective (OIO).

d. The stage is the flat surface with the round opening or aperture in the center. The slide or the specimen
to be viewed is positioned on the stage over the aperture. The stage may be equipped with a mechanical
device into which the slide is clipped. This so-called mechanical stage allows the slide to be manipulated
by means of the two knobs located underneath the stage. Other microscopes do not have a mechanical
stage, and stage clips are used to fasten the slides on the stage.

e. The condenser is a lens located under the stage aperture, and its distance from the stage is controlled by
a knob. Its function is to condense the light and focus it on the specimen.

f. The iris diaphragm is beneath the condenser and functions to control the amount of light passing
through the specimen. This can be moved from side to side to affect a change in light intensity.

g. The illuminator is the source of light which illuminates the object being viewed. This may be replaced
by a mirror.

h. The illuminator control knob, located on the side of the base just below the mechanical stage
adjustment knobs, controls the light output of the illuminator.

i. The adjustment knobs are located on both sides of the microscopes, just above the base. There are two
of these, one located outside the other.
• The coarse adjustment knob is the bigger of the two. Its function is to focus using the scanning
and low power objectives.
• The fine adjustment knob is used in focusing at the high-power and oil immersion objectives.

B. Magnification.

The compound microscope combines the magnifying power of the eyepiece with that of the objective lens.
The magnifying power is marked on the housing of each lens. Simply multiply the magnification values of
the objective and the eyepiece to see how many times the specimen is enlarged. Calculate the total
magnification of the microscope with each objective. Write the answer on the Activity Sheet.

C. Resolving Power.

This is a measure of lens quality. Quality lenses have high resolving power, the ability to deliver a clear
image in fine detail. If the magnification of the lens is high, but the resolution is low, it is of little value.
Although such an image would be large, it would not be clear enough to show details. Resolution is also
affected by the cleanliness of the lens. It is a good practice to clean the lenses with lens paper before and
after every use.

D. Field of View.
This is the size of the area that the lens views. The higher the magnifying power of an objective lens, the
smaller the area viewed. When one switches to a lens with higher magnification, the central portion of what
was visible under low power is seen. It is important to center the specimen before increasing magnification,
to prevent one from losing sight of the specimen.

E. Using the Microscope

1. Cut a small letter "e" from a newspaper article and mount it on a clean glass slide. Add a drop of water
before placing a coverslip. Be sure that no bubbles are formed.
2. Open the spring arm of the mechanical stage, place the slide in the proper position on the stage and
carefully return the spring arm to its closed position. This will hold the slide in place. Using the control
knobs, maneuver the slide on the stage and finally position the tiny "e" over the stage aperture.

3. Illuminate the field of view by maneuvering the mirror. Turn the scanning objective so that it snaps into
place directly over the aperture. Now raise the stage so that the slide is as close to the objective as possible.
While viewing through the eyepiece, use the coarse adjustment knob to move the slide away from the
objective and bring the letter "e" into focus. Using the mechanical stage, move the "e" around. If the slide
is moved to the right, what is the apparent direction of movement of letter "e"? Look at the tiny "e" with
the naked eye and sketch a picture of it. Now look at it through the microscope and sketch that image. Is
there a difference? Carefully center the "e" in the field and proceed to the next step.

4. Without making any other adjustment to the microscope, turn the low power objective into place. Focus
the image using coarse adjustment screw and compare it to the previous one. Is it still in the center of the
field? If not, center it once more.

5. Again, without making other adjustments, rotate the high power objective into place. Viewing the
objective from the side, note how close it is to the slide. Look through the eyepiece, and, using the fine
adjustment knob, bring the "e" into focus. If all is dark, move the slide slightly. Now use the iris diaphragm
to change the amount of light. Did it make a difference?

6. A feature of a good microscope is its parfocal capability. This means that when a specimen is in focus
under low-power magnification, one can switch to high-power magnification and have the specimen remain
in good focus. Usually, a slight adjustment with the fine focus knob is all that is required for a sharp image.

7. After focusing letter “e”, turn the scanning objective back into place and remove the slide.

F. The Dissecting Microscope

The dissecting microscope is useful when magnifications between 5x an50x are desired. It is useful for
viewing entire specimens of small animals or body parts of larger animals. It has the added advantage of
giving a three-dimensional view of the object.

All precautions and rules that apply to the compound microscope also apply to the dissecting microscope.
Some dissecting microscopes have built-in light sources; others do not.

1. Obtain a dissecting microscope. Note that the microscope is binocular and has a single focusing knob
and lens system that allows the change magnification by rotating a dial or revolving nosepiece.

2. Get a 1-peso coin and view this under

the dissecting microscope. Adjust the eyepieces
to fit the width of the eyes. Examine your
specimen under low and high magnifications.
Note the three-dimensional image. Compare the
orientation of the image to the orientation of the
specimen on the stage. Move
the specimen to the right and left.

3. The stage of the microscope is usually equipped with a plate that can be reversed to provide either a
white or black background. Use the background that will provide better contrast. Try switching back and
forth.
Living and preserved specimens are best viewed while completely immersed in water. This prevents
formation of distorted image due to the refraction of light off moist surfaces.

Surnames: ________________________________________________ Group No.: ______________

Course/Section: ____________________________________ Date: ___________________

LABORATORY ACTIVITY 1
THE MICROSCOPE

A. Parts of the microscope. Draw and label the parts of the microscope.
B. Give the function of the following parts of the microscope.

Microscope Parts Function

1. eyepiece or ocular The specimen is viewed through this lens and has
a 10x magnification.
2. revolving nosepiece “Revolves” on 3 magnifications, namely scanner,
low-power objective, and high-power objective.
Some feature the oil-immersion objective.
3. fine adjustment knob Provides small adjustments and focuses on HPO
and OIO.
 4. coarse adjustment knob Provides large-scale zooming adjustments,
focusing mainly on LPO and scanner.
5. arm Connects the body tube and the base, and helps
the viewer hold the microscope carefully.
6. condenser Focuses the light on the specimen.
7. iris diaphragm Adjusts the amount of light passing through the
specimen.
8. scanner Zooms the microscope by 4x.
9. low power objective Zooms the microscope by 10x.
10. high power objective Zooms the microscope by 40x.
11. oil immersion objective Zooms the microscope by 100x.
12. stage Is where the specimen is held. Has an aperture or
a circle where the specimen can be seen.
13. stage clip Fastens the specimen
14. illuminator Is the source of light of the microscope.
15. illuminator control knob Adjusts the brightness of the light source.

C. Magnification

Eyepiece x Objective Total Magnification

Scanner ___10_____ ____4____ ________40_________
LPO ____10____ ____10____ ________100_________
HPO ____10____ ____40____ ________400_________
OIO ____10____ ____100____ _________1000________

D. Draw letter “e” seen under the microscope.

LPO HPO
E. Draw one-peso coin under the dissecting microscope
 Magnification: ___________ Magnification: ___________

F. Guide questions:

1. List down at least three guidelines on how to properly handle and use the microscope.
- Hold it with two arms, placing one on the arm and one beneath the base.
- Start with scanner to locate the specimen and locate its center.
- Clean the lenses before and after use to properly view the specimen.

2. Why must one center and focus the object on the field of view under low power objective before
switching to high power objective?

Because high power objective is for better magnification and detailed view of the specimen,
while lower magnification such as the low power objective and scanner are used to locate and
focus on the specimen.

3. Differentiate magnification and resolution.

Magnification – How large the specimen would appear through the lens.
Resolution – How clear and detailed the specimen would appear through the lens.

4. Compare the image seen in the compound microscope and dissecting microscope in terms of
dimensions, inversion of image, and direction of movement.

Compound microscope – Two-dimensional, inverted image, opposite direction of movement.

Dissecting microscope – Three-dimensional, upright, same movement.

Documentation: Here's a short narration of how the experiment was done:

- A small letter “e” was cut from manila paper, mounted on a slide with water, and covered with a
coverslip.

- The slide was placed on the mechanical stage, and the scanning objective was used to locate
the specimen.
 - The coarse adjustment knob focused the image, revealing its inverted movement.
- The letter was sketched, then refocused and recentered under LPO and HPO using the fine
adjustment knob.
- The iris diaphragm was adjusted for better lighting, and the slide was removed after
observations.
- A 1-peso coin was examined under a dissecting microscope, appearing three-dimensional and
upright.
- The image moved in the same direction as the specimen, and different backgrounds were
tested for contrast.
- The experiment highlighted differences in image orientation, magnification, and depth
between the two microscopes.

References: