Exercise 2

MICROSCOPY

After completing this exercise, the student should be able to:

1. Know the different parts and respective functions of the

compound microscope;
2. Use the microscope effectively, especially the oil-
immersion lenses;
3. Manipulate the compound microscope proficiently;
4. Handle and care for the compound microscope properly.

MATERIALS

Yeast suspension of Saccharomyces cerevisiae

Glass slides
Cover slips
Methylene blue staining solution
Droppers

PROCEDURE

1. Examine the microscope and locate the components listed

below:

a. Ocular h. Condenser
b. Body tube (or barrel) i. Iris-diaphragm lever
c. Revolving nosepiece j. Coarse adjustment knob
d. Low power objective k. Fine adjustment knob
e. High power objective l. Condenser-adjustment knob
f. Oil immersion lens m. Mechanical stage
g. Stage n. Base

2. The microscope should have been stored with the low power
objective in position. If such is not the case turn the
nosepiece with the low power (10x) objective clicks into
position and make a mental note always to return it to that
position for storage. Look through the ocular lens while
adjusting the mirror to direct 1 cm above the stage, with the
condenser racked up to full top position, and with the
diaphragm half open. With the mirror in position observe the
effects of lowering the condenser and changing the iris
diaphragm opening.

3. With a dropper place a drop of water near the center of a

clean slide and stir into a small amount of yeast suspension
 picked up with your inoculating loop. Avoid too dense
suspension as the mixture should appear only faintly turbid
on the slide. Place a cover slip over the suspension,
position the slide on the stage with the area to be examined
directly over the condenser lens and examine with the
microscope at low power.

4. Lower the objective almost to the surface of the slide and

focus upward with the course adjustment until the yeast
preparation is approximately in focus. Then use the fine
adjustment wheel for sharp focus. Rotate the eyepiece to
indicate the presence of extraneous material on the lens
which appears to be a part of the microscopic field. If the
ocular is dusty, have your instructor examine it. Record your
observations and prepare sketches of the yeast cells.

5. Alter the light condenser position and the slide position,

making note of the effects of three manipulations.

6. Examine a suitable field at higher magnification. First

elevate the body tube to raise the nosepiece and turn the
high power objective into position. For all objectives other
than low power the condenser should be elevated to the full
top position. Again reach a rough focus by elevating the
draw tube from a too close position and switch to the fine
adjustment. Adjust the diaphragm for proper illumination and
record observations at this magnification.

7. Repeat with the oil immersion lens, but take great care to
avoid contact between lens and cover slip. The working
distance is very short and the following procedure is
required. Add one drop of immersion oil to the cover slip;
while watching from the side lower the lens carefully until
it makes contact with the oil, then focus upward with the
fine adjustment until the field comes into focus. Have the
iris diaphragm more open than for the high power objective.
Note budding cells and granules within the yeast cells.

8. Prepare a second slide with yeast cells suspended as before,

but allow this suspension to air dry thoroughly without cover
slip. Cover the dried film with methylene blue for a minute,
gently rinse off the dye with tap water and air dry. Note
the appearance of the stained yeast preparation at each
magnification without the use of a cover slip.
QUESTIONS

1. a) What is the total magnification obtainable with the lower

power objective?

b) What is the total magnification obtainable with the oil

immersion lens?

5. Complete the following table:

MICROSCOPE PART FUNCTION(S)

Ocular

Objective

Condenser

Iris-diaphragm lens

Fine adjustment knob

Coarse adjustment knob

Condenser adjustment
knob
Base
Arm

Stage

Stage Clips

Mirror
6. Identify & label the parts of a compound microscope.
7. Give 5 practices/ways on how to handle and take care for
the microscope properly.