Laboratory Activity 1.

Microscopy: The Compound Microscope

In biology, it is imperative that the student must be proficient in the use of the microscope.
A microscope is an instrument that uses lenses to produce an enlarged, focused image of a
specimen. It is used to study cells, tissues, and minute organisms (i.e., protozoans, bacteria,
viruses, etc.) that are too small to be seen by the naked eye. Basically, there are two types of
microscopes, the light microscope focuses light onto a specimen using a mirror or a light bulb, and
the electron microscope uses a beam of electrons to create an image.
Ideally, at the conclusion of this exercise, you should be well on your way to success in the
use of the microscope. From this point on to the succeeding laboratory activities that require
microscopic examinations, the compound light microscope shall be used.

A. Mechanical Parts

1. Base- usually the horseshoe pad that supports the entire microscope; provided with
contact pads on the underside to give stable support on surfaces, balance, and stability
when used in an inclined position.
2. Pillar- the slightly elevated part that connects the base to the arm and supports the
upper part of the microscope.
3. Arm- the curved part that one holds in carrying the microscope.
4. Inclination joint- a movable hinge that uses in tilting or inclining the microscope in the
desired position.
5. Body tube- cylindrical hollow tube attached to the arm and serves as a passageway of
light from primary and secondary magnifiers.
6. Draw tube- a smaller cylinder at the upper part of the body tube that holds the eyepiece.
7. Revolving nosepiece- a rotating head attached to the base of the body tube which
carries and facilitates the shifting of the objectives.
8. Dust shield- a fixed circular disc above the nosepiece that protects the objective lens
from dust.
9. Stage- a square or rectangular plate with a flat surface on which the specimen rest.
10. Sub-stage- a structure found below the stage that holds the condenser and iris
diaphragm.
11. Coarse Adjustment Knob- a larger pair of knobs used when focusing the specimen with
low power objective by raising and lowering the body tube.
12. Fine Adjustment Knob- a smaller pair of knobs used when finally focusing the specimen
with either low-power or high-power objectives.

B. Illuminating Parts

1. Mirror- usually with a double-sided face with a concave and plain surface, used when
the light source is at low and very bright intensity, respectively.
2. Condenser- a simple optical unit that fills the field of view with uniform illumination and
supplies the objective with a cone of light at the right size to produce maximum results.
3. Iris Diaphragm- attached at the base of the sub-stage ad provided with a lever or
projecting handle. This can be manipulated to regulate the amount of light passing
through the lenses.

C. Magnifying Parts

1. Eyepiece- also known as ocular

- An optical unit that is usually in the form of a detachable cylinder provided
with lenses found above the draw tube.
2. Objectives- produces the magnified image of the object that shows all detail structure
and general appearance from which all observations are made. Generally, a compound
light microscope is provided with three objectives, namely:
a. Low Power Objective (LPO)- the shortest cylinder with a large lens opening and has
lower magnification. Used to obtain a large field of magnification thus, observe the
general view of the specimen.
b. High Power Objective (HPO)- with a longer cylinder and smaller lens opening than
LPO. Used to obtain the detailed parts of the specimen with higher magnification.
 c. Oil immersion Objective (OIO)- the longest cylinder with a small opening from other
objectives attached in the microscope. Used to observe fine details of specific parts
with a very high magnification.

Proper Care and Handling of the Microscope

The objectives (lenses) of the microscope and the ocular (eyepiece) must be kept clean or
the image will be distorted. The lenses should be cleaned periodically, but only with lens paper. If
you see obvious dirt, blurred specks, or similar things which disturb the field of vision, something
needs cleaning. If the specks move when you rotate the ocular, this usually means that the ocular
needs cleaning. If they do not remove when the ocular is rotated, either the objective needs
cleaning or the cover glass of the slide is dirty. If it is the cover glass, the dirt will move when the
slide is moved. If an objective needs cleaning, do the job without removing it from the mount. If
nothing seems to work, ask your instructor for assistance. Do not take anything apart.

Follow your instructor’s directions on the use of a specific, assigned microscope. When
carrying your microscope to and from the cabinet, carry it upright (vertically) by the arm, with one
hand under the base. When returning the microscope, make absolutely sure that you have not left
a slide on the stage.

Objectives
This activity will guide the students on the proper use and handling of a compound
microscope. Students should also identify the basic parts and functions of a compound
microscope.

Material

Compound microscope, glass slide, coverslip, water, medicine dropper, cut-out of the smallest
letter “e” from newspaper or magazine.

Procedure

1. Place the microscope on the table with the arm facing you.
2. Identify the different parts of a compound microscope by drawing and labelling the parts of
a microscope on the space provided in page 3.
3. Mount the cut-out small letter “e” in an upright position to a glass slide with water. (Figure 1)
Avoid formation of bubbles in the fresh mount preparations so as not to interfere during
observations.
4. Place the mounted letter “e” on the stage and clamp the stage clip.
5. Align the low power objective with the aperture of the stage.
6. While looking at the side of the microscope, lower the body tube by turning the coarse
adjustment knob clockwise until the objective lenses and the specimen stage are about half
an inch. Turning the coarse adjustment knob counter-clockwise will increase the distance
between the objective lens and the stage.
7. Look through the ocular with your “master eye”. Adjust the light from the source toward the
stage aperture. Regulate the light by opening or closing the diaphragm. The field of view
must be evenly lighted.
8. Carefully raise the objective by turning the coarse adjustment knob counter clock-wise until
the specimen comes into clear and sharp focus. If you cannot see the image yet, adjust the
slide and refocus.
9. To obtain a more sharply defined focus, turn the fine adjustment knob slightly clockwise
and counterclockwise.
10. Once the specimen is closely focused, observe the image of the letter “e” as seen under
the microscope. Draw your observations.
11. Carefully, move the specimen slightly upward, downward, and into the left and right .
direction. Note the movement of the specimen.
12. Adjust the position of the letter “e” at the center of the microscope field. Shift to the HPO by

the image seen. e

 rotating the revolving nosepiece until a feeble click sound was heard. Make a slight
adjustment with a fine adjustment knob until a sharp is achieved. Observe and draw the
image seen.

Figure 1. Mounting of letter “e” in upright position at the center of the glass slide.

Guide Questions:
1. What does working distance mean?
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2. What does microscopic field mean?
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3. Describe the letter “e” seen under the microscope? what do you think is/are the reasons for
your observation?
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4. What is the direction of the images seen under the microscope when moved in different
directions?
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5. How far off from the center is the object in your microscope when shifted from LPO to
HPO?
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6. Why is it necessary to use both eyes when observing under the microscope?
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7. What is meant by master eye?

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8. Differentiate compound microscope from electron microscope. What are the advantages
and disadvantages of the use of each microscope?
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9. Write in a short paragraph how would you set-up and focus a specimen using a compound
microscope. Be sure to describe the correct order in using the different focusing dials.
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COMPUTE THE TOTAL MAGNIFICATION
OBJECTIVE OCULAR TOTAL MAGNIFICATION
Scanner
Low power objective (LPO)
High power objective (HPO)
OBSERVATIONS:

Letter “e” as it appears through the eyepiece.

Magnification: _____________

Magnification: _____________