EXERCISE 1

THE COMPOUND MICROSCOPE

I. INTRODUCTION
The microscope is an indispensable tool in Botany as in all Biological Sciences. It enables the user to see
structures too small to be seen by the unaided eye. Since the invention of the light microscope in the
1590s, there have been numerous improvements and modifications on the performance of the
instrument itself. However, part of this improvement depends on the development of new techniques
for specimen preparation. Regardless of the kind of microscope being used, three elements are needed
to form an image: a source of illumination, the specimen to be observed, and a system of lenses to focus
the illumination on the specimen and to form the image. The compound light microscope is a
microscope that uses natural or artificial light and series of lenses: the condenser, the objectives, the
intermediate lenses and the ocular lens. The objective lens magnifies and projects virtual image into the
body tube while the ocular lens magnifies that image further and projects the enlarged image into the
eye.

II. OBJECTIVES

At the end of the exercise, the student is expected to be able to:

1. To identify the different parts and functions of the microscope.

2. To develop basic skills in the operation of the microscope.
3. To determine magnification/reduction in size of objects viewed under the microscope.
4. To become acquainted with the preparation of wet or fresh mounts for microscopic
examination.

III. MATERIALS

Equipment/Apparatus:
Compound light microscopes

Glasswares:
Slides and cover slips

Plant Specimens:
Leaves
Rice grains
Hydrilla verticillata (digman)

Other materials:
Small letter ’e’ (newsprint)
Lens paper or cotton with alcohol
Pair of scissors
IV. PROCEDURE

A. Familiarization with Parts 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 Function/s
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. This is no longer included in the design of current
microscopes.

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 that protects 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 for adjusting the focus of the scanner and low power objective
b. Fine Adjustment Screw for delicate focusing of 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 found below the stage and attached to the pillar; holds the mirror in
place.

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.

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 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 cedar 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 from the cabinet. Use both hands to carry the microscope and hold it in an
upright position. With one hand, secure the microscope by its arm and support the base of the
microscope with your other hand.

2. Set the microscope down in a side view position to better locate the other parts.

3. Identify the parts of your microscope and label the drawing in worksheet 1.

B. Use of the microscope

After familiarizing yourself with the parts of your microscope, you are now ready to use the microscope.

1. Set the microscope down with the arm towards you and the stage away from you.

2. Rotate the nosepiece until the low power objective clicks into position.

3. Slowly raise the stage or lower the objective by rotating the coarse adjustment until the low
power objective is about 1 cm from the stage. When trying to locate the specimen, the low
power objective is always used first before shifting to the high power objective for a detailed
study of the specimen.

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 4. Always relax and keep both eyes open when using the microscope to prevent eye strain.

5. While looking through the ocular, orient the mirror toward a light source so that it reflects the
light up through the open diaphragm, condenser, opening in the stage and body tube. If this is
done properly, a bright, evenly distributed circle of light called the microscope field will be
visible. If the field is too bright, close the diaphragm to reduce the amount of light reflected
from the mirror or if it is too dim, open the diaphragm.

6. Cut a small letter ‘e’ from newspaper clippings available in the laboratory. Place the letter
‘e’ on a glass slide. Cover with a cover slip. Place the slide on the center of the stage.

7. When proper illumination of the field has been achieved, slowly raise the body tube (or lower
the stage) by means of the coarse adjustment knob until the letter comes into focus. It is
helpful to move the slide slowly back and forth while doing this in order to help locate the
materials being studied.

8. Observe the letter under low power. Make a sketch of what you observe in worksheet 1.

C. Preparation of a wet mount

In Botany, you will be asked to make a wet or a fresh mount to facilitate viewing of some plant parts.

1. Use forceps or tweezers to pick off a young leaf from a portion of the digman stem.

2. Place the leaf on the center of a clean glass slide.

3. With a dropper, put a drop of water over the leaf.

4. Get a cover slip and put one edge of the cover slip on one side of the drop of water. Then with a
dissecting needle, slowly bring the cover slip down on the water. If air bubbles form, gently tap
the cover glass. Bubbles interfere with viewing of the specimen.

5. Focus under the LPO of the microscope. Do you see many green organelles? Locate the orange
brown tooth cells along the margin of the leaf. How many are they? Shift to HPO to see more
details of the cells.
6. If you are asked to make wet mounts of roots, stems, or fruits, take a sharp blade and make thin
sections of the specimen. Place these sections on a glass slide and proceed as above in number
4.

D. Computation of magnification

Magnification may be defined as the number of times the image of an object is enlarged by the
magnifying lens or the number of times a drawing is enlarged or reduced from the original size of
the object.

1. To determine the total magnification of the specimen as seen through the microscope, multiply
the magnification of the ocular by the magnification of the objective.

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 Total Magnification = Magnification of eyepiece x Magnification of objective

2. To determine the magnification in a drawing made, divide the size of drawing by the actual
size of object or specimen.

Magnification of drawing = Size of drawing

Actual size of object

3. Compute for the magnification of the length of a potted plant, length of rice grain and diameter
of tooth cell drawn 5 cm long.

4. Show your computation on the worksheet.

E. Reminders on the care of the microscope

1. Never touch the lens of the microscope with your hands. If the lenses are dirty, use lens paper
or cotton with alcohol.

2. If the microscope is out of order, notify your laboratory instructor.

3. Report any missing parts of your microscope.

Exercise adapted from:

De Jesus J, Ragragio E. 2000. Laboratory Manual in General Botany. De La Salle University Press, Inc.,
Manila. pp. 1-9.

Other References:
Abramoff P, Thomsom R. 1982. Laboratory Outlines in Biology III. W. H. Freeman and Co. San
Francisco. pp. 1-9.
Barbour MG, Bonner BA, Breckon GJ. 1975. Botany: A Laboratory Manual. 5th edition. John Wiley &
Sons, Inc. New York. pp. 5-10.
Becker WM, Reese JB, Poenie MF. 1996. The World of the Cell. The Benjamin/Cummings
Publishing Company, Inc. California. pp. 841-845.