EXERCISE 1

THE COMPOUND MICROSCOPE

1. Introduction
3. To become familiar with measuring
The microscope is an indispensable tool microscopic objects.
in zoology and in all Biological Sciences.
It enables us to see animal structures 4. To determine magnification/reduction
that are too small to be seen by the in size of objects viewed under the
unaided eye. Since the invention of the microscope.
light microscope in the 1590’s, there
have been numerous improvements and 5. To become acquainted with the
modifications on the performance of the preparation of wet or fresh mounts
instrument itself. However, part of this for microscopic examination.
improvement depends on the
development of new techniques for III. Materials
specimen preparation. Regardless of
the kind of microscope being used, Equipment/Apparatus:
three elements are needed to form an Compound microscopes
image: a source of illumination, the Ocular and stage micrometers
specimen to be observed, and a system Prepared slide of a blood smear
of lenses to focus the illumination on the
specimen and to form the image. The Glasswares:
compound light microscope is a Slides and cover slips
microscope that uses natural or artificial
light and series of lenses: the Stain:
condenser, the objectives, the Lugols Iodine
intermediate lenses and the ocular lens.
The objective lens magnifies and Specimens:
projects virtual image into the body tube Cheek cells
while the ocular lens magnifies that
image further and projects the enlarged Other materials:
image into the eye. Small letter ’e’ (newsprint)
Lens paper or cotton with alcohol
II. Objectives Immersion oil
Pair of scissors
At the end of the exercise, we will be Tooth Pick
able to: Ruler

1. To identify the different parts and 1V. Procedure

functions of the microscope.
A. Familiarization with Parts of the
2. To develop basic skills in the Microscope
operation of the microscope.
The three basic parts of the microscope
are: (a) mechanical parts; (b) magnifying
parts; and (c) illuminating parts.
A. Mechanical Parts (i.e. those parts concerned with the support and adjustment of the optical
parts)

Parts Functions
1. Base Stand that supports the microscope.
2. Pillar A short piece of metal that attaches to one end of the
base; also supports the microscope.
3. Handle or Arm Curved metallic part arising from the pillar used for holding the
microscope.
4. Inclination Screw Found at the junction of the pillar and the handle used for tilting
the microscope.
5. Body Tube Cylindrical structure vertically arising from the handle; holds the
dust shield and nosepiece.
6. Ocular Tube or Draw Tube Upper smaller end of the body tube bearing the eyepiece or
ocular lens.
7. Revolving Nosepiece Circular structure where the objectives are attached that permits
the shifting of objectives.
8. Dust Shield Circular structure above the nosepiece used to protect the
lenses of the objectives.
9. Adjustment Screws Two pairs of wheels attached to either side of the body tube.
These are:
a. Coarse Adjustment Screw Used to adjust the low power objective in focusing.
b. Fine Adjustment Screw Used for delicate focusing in connection with the high power and
oil immersion objectives.
10. Stage A square or round platform with an opening at the center where
the slide is placed.
11. Mirror Rack/light aperture Found below the stage and attached to the pillar; holds the
mirror in place.
12. Stage adjustment knob Moves the specimen slide forward, backward and sideways

B. The Magnifying Parts (i.e. those parts concerned with image enlargement of the specimen)
Parts Functions
1. Ocular or Eyepiece Found on the draw tube through which the operator peeps
during actual focusing; usually carries magnification of 10X.
A thin, black line that appears to cut halfway across the field of
view which the student or instructor can use to point out regions
of the specimen under observation is called a pointer.
2. Objectives : Tube-like structures attached to the revolving nosepiece.
These are:
a. Scanner Carries a magnification of 4-5X.
b. Low Power Objective (LPO) Carries a magnification of 10-12X.
c. High Power Objective (HPO) Carries a magnification of 40-60X.
d. Oil Immersion Objective Carries a magnification of 97-100x; requires the use of
immersion oil.
C. The Illuminating Parts (i.e. those parts concerned with light provision and regulation to the
specimen.
Parts Functions
1. Mirror Found below the stage near the base used to collect and direct light to the
specimen.
2. Diaphragm Found below the stage used to regulate the entry of light onto the specimen.
There are different types of diaphragm: iris, plate or fan.
3. Condenser Lens found immediately beneath the hole of the stage used to concentrate
light rays on the specimen.
1. Get a microscope. Use both hands to amount of light reflected from the
carry the microscope and hold it in an mirror or if it is too dim, open the
upright position. With one hand, diaphragm.
secure the microscope by its arm and
support the base of the microscope 6. Cut a small letter ‘e’ from newspaper
with your other hand. clippings available in the laboratory.
Place the letter ‘e’ on a glass slide.
2. Set the microscope down in a side Cover with a cover slip. Place the
view position to better locate the slide on the center of the stage.
other parts.
7. When proper illumination of the field
3. Identify the parts of your microscope has been achieved, slowly raise the
and label the drawing in worksheet 1. body tube (or lower the stage) by
means of the coarse adjustment knob
B. Use of the microscope until the letter comes into focus. It is
helpful to move the slide slowly
After familiarizing yourself with the parts back and forth while doing this in
of your microscope, you are now ready order to help locate the materials
to use the microscope. being studied.

1. Set the microscope down with the 8. Observe the letter under low power.
arm towards you and the stage away Make a sketch of what you observe in
from you. worksheet 1.

2. Rotate the nosepiece until the low C. Measurement of microscopic

power objective clicks into position. objects using a micrometer
eyepiece
3. Slowly raise the stage or lower the
objective by rotating the coarse Sometimes the biologist needs to know
adjustment until the low power the dimensions of the object being
objective is about 1 cm from the observed under the microscope. If you
stage. When trying to locate the want to estimate the size of microscopic
specimen, the low power objective is objects more accurately, a microscope
always used first before shifting to the equipped with an ocular micrometer can
high power objective for a detailed be used.
study of the specimen.
An ocular micrometer is a small glass
4. Always relax and keep both eyes disc etched with uniformly spaced lines.
open when using the microscope to When using an ocular micrometer in
prevent eye strain. examining an object under a
microscope, the micrometer acts as a
5. While looking through the ocular, ruler that is superimposed on the object
orient the mirror toward a light being measured. However, the units of
source so that it reflects the light distance between the spaces of the
up through the open diaphragm, ocular micrometer are arbitrary.
condenser, opening in the stage and Therefore, before an ocular micrometer
body tube. If this is done properly, a can be used for actual measurements of
bright, evenly distributed circle of microscopic objects, it must first be
light called the microscope field will calibrated against a standard. You need
be visible. If the field is too bright, a stage micrometer for calibration.
close the diaphragm to reduce the
A stage micrometer is a glass slide with a given number of spaces in the
scribed lines that are exactly 0.01 mm ocular micrometer.
(or 10 micrometers) apart.
8. Since the smallest spaces on the
Calibration of an ocular micrometer stage micrometer are known to be
0.01 mm apart, compute the
1. Get an ocular and a stage distance of 1 ocular division or
micrometer. These are delicate calibration constant as follows:
instruments and must be protected
from dust, grease, fumes, and
friction. These micrometers must be
handled at the edges.

2. How to attach an ocular micrometer Where: od = ocular division

to the microscope. ms = micrometer spaces

Detach the ocular tube from the 9. What is the calibration constant of the
draw tube. microscope you are using? Show
your computation on the worksheet.
Unscrew the eyepiece lens (the top
lens for some microscopes but for 10. You may now remove the stage
Zeiss microscopes, it is the lower micrometer and replace it with the
part of the eyepiece) of the ocular slide containing the specimen that
tube. Very carefully insert the ocular you want to measure.
micrometer.
11. The calibration constant is used to
Put back the eyepiece lens and determine the length or width of the
insert the ocular tube back into the microscopic specimen that you are
draw tube. measuring. Multiply the number of
divisions of the micrometer eyepiece
3. Mount the stage micrometer. Find covered by the specimen by the
the center of the scale and focus calibration constant.
with the low power objective.
Length or Width = No. of ocular ms x calibration
constant
4. While looking through the low power
objective, rotate the ocular and
D. Use of the Oil Immersion Objective
adjust the stage micrometer until
(OIO)
the lines of the ocular micrometer
are parallel with those of the stage
1. Obtain a prepared slide of a blood
micrometer.
smear and focus under LPO.
5. Match the lines at the left edges of
2. When a clear image has been
the two micrometers by moving the
obtained, turn the revolving nose piece
stage micrometer.
to a higher magnification (HPO) until
you obtain a clear image.
6. Observe another point where the
lines of the two micrometers
3. Obtain immersion oil provided by the
coincide again.
laboratory technician. Put a drop of oil
7. Now count the number of spaces of
on top of the coverslip of the specimen.
the stage micrometer included within
Rotate the objective from HPO to OIO.
DO NOT USE THE COARSE
ADJUSTMENT KNOB AT THIS TIME. F. Computation of magnification
Use only the fine adjustment knob to
obtain a clear image of the specimen. Magnification may be defined as the
number of times an object is enlarged
4. Locate a red blood corpuscle (RBC) by the magnifying lens or the number of
and any of the white blood corpuscles times a drawing is enlarged or reduced
(WBC) in the smear. from the original size of the object.

5. Measure the diameter of 10 RBC and 1. To determine the total magnification

3 WBC and get the average of each. of the specimen as seen through the
Show your computation in your microscope, multiply the magnification
worksheet. of the ocular by the magnification of the
objective.
6. Once you are done using the OIO,
clean the glass slide by wiping with Total Magnification = Magnification of eyepiece x
Magnification of objective
clean cotton dipped in 70% alcohol.
Clean the lens using lens paper only.
2. To determine the magnification in a
drawing made, divide the size of
E. Preparation of a wet mount
drawing by the actual size of
object or specimen.
1. Prepare a wet mount of cheek cells.
Using a toothpick, gently scrape off Magnification of drawing = Size of drawing
the inner walls of your cheek and Actual size of object
smear it on the center of a clean
microscope slide. 3. Draw a 5 cm diameter cheek cell in
2. Add a drop of Lugol’s iodine on the your worksheet. Compute for the
smear. Get a cover slip and put one total magnification of the drawing.
edge of the cover slip on one side of
the drop of stain. Then with a 4. Show your computation on the
dissecting needle, slowly bring the worksheet.
cover slip down on the drop of stain.
If air bubbles form, gently tap the
cover glass. Bubbles interfere with G. Reminders on the care of the
viewing of the specimen. microscope

3. Focus under the LPO of the 1. Never touch the lens of the
microscope. Observe the general microscope with your hands. If the
structure of the cell. Locate the lenses are dirty, use lens paper in
basic parts of the cells (cell cleaning the lens of the objectives
membrane, cytoplasm and nucleus). or cotton with alcohol for cleaning
Measure the length and diameter of the OIO.
the cell. What is the length and
diameter of the cheek cell? Show 2. If the microscope is out of order,
your computation on the worksheet. notify your laboratory instructor.

3. Report any missing parts of your

microscope.
References

1. Abramoff, P. and R. Thomsom. 1982. Laboratory Outlines in Biology III. W. H.

Freeman and Co. San Francisco. pp. 1-9.

2. Barbour, M. G., B. A. Bonner and G. J. Breckon. 1975. Botany: A Laboratory

Manual. 5th edition. John Wiley & Sons, Inc. New York. pp. 5-10.

3. Becker, W. M., J. B. Reese and M. F. Poenie. 1996. The World of the Cell. The
Benjamin/Cummings Publishing Company, Inc. California. Pp. 841-845.
Student’s Name ___________________________Date Performed __________
Course/Year/Section _______________________Date Submitted __________
Professor’s Name___________________

WORKSHEET # 1

THE COMPOUND MICROSCOPE

A. Label the parts of the microscope

B. Using the microscope

Before viewing Viewed under the microscope

Sketch of the letter e using the LPO. Sketch of the letter e using the HPO

1. Is the position of the letter e the same as when viewed with the naked eye
or is it inverted?

2. When you move the slide to the left, in which direction did the letter image
appear to move?

3. When you move the slide toward you, in which direction did the letter image
appear to move?

4. Explain your observations.

5. When you shift to HPO, what happens to the letter image and field of view?
C. Measurement of microscopic objects using a micrometer eyepiece

Calibration constant (computation)

Width/length of the cheek cell

1 cm

E. Magnification

Specimen Actual size Magnification of a 5cm

drawing
(Show computation)

Cheek cell (diameter)

WBC (diameter)

RBC (diameter)

Frog (crown-rump length)

10 cm
Label the parts of this light microscope.
The parts and function of the stereomicrscope is basically similar with that of the compound light
microscope. Using your idea of the parts and function of the compound microscope label the parts of this
stereomicrscope by inferring on its functions. Manipulate and explore the use the microscope for you to
have an idea regarding the parts and function of this microscope.