1

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.
Objectives At the end of the activity, the student
should be able to
1.

THE
MICROSCOPE

2.
3.

identify the parts of a typical microscope and

learn their functions
focus
specimens
under
the
compound
microscope and dissecting microscope
measure specimens under the microscope

Materials
brightfield compound microscope
dissecting microscope
newspaper cut-out
glass slides

one peso coin

ocular micrometer
stage micrometer
coverslips

Procedure
A. The Microscope and its Parts
Locate the parts of the microscope listed below and
label the illustration on the Activity Sheet:
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 focused. 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 oilimmersion 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 substage 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 substage 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 oilimmersion 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?

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 fairly good focus. Usually a slight
adjustment with the fine focus knob is all that is required for a sharp image.
6.

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

F.

Measuring Specimen Using Ocular Micrometer

More precise measurements can be made using an ocular micrometer. An ocular

micrometer is a thin circle of glass or plastic etched with a non-unit scale usually
ranging from 0 to 100. When placed in the ocular lens of the compound microscope,
the scale image is superimposed over the image of the specimen. A stage
micrometer is a glass slide etched with a scale of known units, usually divided into
0.01 mm graduations.
The ocular micrometer is calibrated using the stage micrometer by aligning the
images at the left of the scales (figure 1). To calibrate, do the following:
1.
2.
3.
4.
5.
6.
7.

Install the 10x ocular containing the ocular micrometer disc in the microscope.
Place the calibrated stage micrometer slide on the stage and focus on the scale.
Adjust the field so that the zero line of the ocular disc scale is exactly
superimposed upon the zero line of the stage micrometer scale.
Without moving the stage micrometer, locate the first point to the right where
any two lines are exactly superimposed upon each other.
Count the number of divisions or spaces (mm) on the stage micrometer
between the zero line and the superimposed line to the right.
Count the number of ocular divisions or spaces between the zero line and the
superimposed line to the right.
Divide the spaces in the stage micrometer determined in step 5 with ocular
spaces in step 6 and multiply by 0.01mm, the measurement in each division in
the stage micrometer.
1 ocular space /calibration constant (mm)/ =Stage micrometer spaces X 0.01 mm
Ocular micrometer spaces
To express the calibration constant in micrometer ( m), multiply the ocular
space in mm by 1000.

8.

Repeat steps 3 through 7 for each objective on the microscope. If the ocular
micrometer is moved to a different microscope, the calibration procedure must
be repeated. If a new objective is added to the microscope, the calibration
procedure must be done for the objective.

9.

Compute the calibration constant of the microscope under LPO. Show the
computation on the worksheet.

10. Remove the stage micrometer and replace it with the prepared slide of
Paramecium.
11. Use the calibration constant to determine the length and width of Paramecium.
Multiply the number of divisions in the ocular micrometer covered by the
specimen by the calibration constant.
Length or width = number of spaces covered in the ocular micrometer X
calibration constant
12. Record the measurements on the activity sheet.
G.

The Dissecting Microscope

The dissecting microscope is useful when magnifications between 5x and 50x 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 in 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. Write the observations on the Activity Sheet.
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.

References
Todd, JC. Clinical Diagnosis by Laboratory Methods. Philadelphia, PA,
W.B.Saunders Co.
Miller, Stephen A: General Zoology Laboratory Manual. 5th ed. McGraw-Hill.
http://webnt.calhoun.edu/distance/Internet/Natural/Biol103/New/HTML/Bio103/h.tips.
http://www.vscc.cc.tn.us/academic/math/biol/biol100/lab1.htm

Activity #1
THE MICROSCOPE
Student's Name _____________________________ Date Performed ___________
Course/Year/Section _________________________ Date Submitted ___________
Instructor's Name ____________________________
I. A. Label the parts of the microscope
B. Magnification
Eyepiece

Objective

Total Magnification

_______
_______
_______
_______

x
x
x
x

________
________
________
________

=
=
=
=

_______________
_______________
_______________
_______________

Scanner
LPO
HPO
Oil immersion
C. Draw letter "e"

1. normal position
2. under low-power objective
3. under high-power objective
D. The Ocular Micrometer
1. 1. Compute for the calibration constant of your ocular micrometer. Show
your solution.
Under LPO:
Under HPO:
1.2. How do these compare with each other?
2.1. Determine the width (in m) of the Paramecium (same area). Show your
solution.
Using LPO:
Using HPO:
2. 2. How do these compare with each other?

II. Guide Questions

1. What is the correct procedure for carrying the microscope?
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2. Compare the image seen in the compound microscope and dissecting
microscope in terms of magnification, dimensions, inversion of image and
direction of movement. Tabulate your answers

3. How does one improve contrast when using a dissecting microscope?

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