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
examinatio
n.

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 Screw
Screws two pairs of wheels attached to either side of the body tube. These
are: for adjusting the focus of the scanner and low power objective
a. Coarse Adjustment for delicate focusing of the high power and oil immersion
Screw b. Fine Adjustment

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.

A thin, black line that appears

a. Scanner
to cut
b. Low
halfway
Power
across the field of
view which the student orObjective
instructor(LPO)
can use
c. High
to point out regions of
the specimen under observation
Power Objective
is called a(HPO) d.
Oil Immersion Objective
2. Objectives carries a magnification of 4-5X. carries a magnification of 10-
: 12X. carries a magnification of 40-60X. carries a
magnification of 97-100x;requires the use of cedar oil.
e structures attached to the revolving nosepiece. These

C. The Illuminating Parts (i.e. those parts concerned with light provision and regulation
to the
specime
n.

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.

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.

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.

Name________ ____________________________ Date Performed _________________

Biology 1 Section __________________________ Date Submitted __________________
Teacher __________________________________
 WORKSHEET # 1 THE
COMPOUND
MICROSCOPE

A. Label the parts of the

microscope.

Figure 1. The parts of a compound light

microscope
 6

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 does the letter image appear to
move?

3. When you move the slide toward you, in which direction does 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?
 6. What is the total magnification of your microscope when the given objective is in
place?
Scanner ______________ High Power Objective (HPO) _______________
Low Power objective ________ Oil Immersion Objective (OIO) _____________

C.
Magnification

Specimen Actual size Magnification of a 5cm

drawing (Show computation) Hydrilla tooth cell (diameter) 0.01mm

Length of a potted
plant

Rice grain
(length)
8