ACTIVITY NO.

1
THE USE AND CARE OF THE MICROSCOPE

I. INTRODUCTION

The microscope is a very important tool for a microbiologist. Beginning students frequently become
impatient with the microscope and forgo every opportunity to practice and develop their observation
skills. Simple observation is a critical part of science. Making discoveries by observation requires
curiosity and patience. Procedures for observation cannot be provided. Sketches and/or drawings are
required instead to enhance effective observation. Students need not be artists to draw what they see. In
drawing, the following need special attention:
1. Size relationships. For example, how big are bacteria relative to protozoa?
2. Spatial relationships. For example, where is one bacterium in relation to the others? Are they
all together in chains?
3. Behavior. For example, are individual cells moving or are they all flowing in the liquid
medium?
4. Sequence of events. For example, were cells active when you first observed them?

II. OBJECTIVES

After completing this activity, the students should be able to:

1. identify the parts of the microscope and their functions
2. become familiar with the 3 variations of light microscopy
3. learn how to use the microscope effectively particularly the oil immersion objective
4. identify the three basic morphologies of bacteria.

III. MATERIALS
Microscope
Immersion oil
Lens paper
Prepared slides of bacteria (cocci, bacilli, spirilli)

IV. PROCEDURE

1. Place the microscope on the table squarely in front of you. Identify all parts. Draw the
microscope and label all the parts.

2. Obtain a slide of bacteria and place it on the stage.

3. Adjust the eyepieces on the microscope to your own personal measurements.

a. Look through the eyepieces and, using the thumb wheel, adjust the distance between
the eyepieces until one circle of light appears.
b. With the low power (10X) objective (LPO) in place, focus the microscope on the slide
.
4. Raise the condenser up to the stage. On some microscopes, the condenser can be focused by
the following procedure:
a. Focus with the 10X piece.
b. Close the iris diaphragm so only a minimum of light enters the objective lens.
c. Lower the condenser until the light is seen as a circle in the center of the field.
 d. Raise the condenser up to the slide, lower it, and stop when the color on the periphery
changes from pink to blue.
e. Open the iris diaphragm until the light just fills the field.

5. Draw some of the cells on the slide under low power.

6. Shift to the high power by rotating the turret to the next lens. The object of study may be
slightly blurred so that it is necessary to refocus using the fine adjustments.

7. Draw some of the cells under high power.

8. Move the high power objective out of position and place a drop of immersion oil on the area of
the slide in focus. Carefully click the immersion lens into position. It should now be
immersed in oil. Fine adjustment should bring the object into focus.

9. Draw some of the cells under oil immersion objective.

10. When your observations are completed, move the turret to bring a low power objective into
position. DO NOT rotate the high power objective through the immersion oil. Clean the oil
off the objective lens with lens paper, and clean off the slide with tissue paper or a paper towel.
Keep the slide.
 LABORATORY REPORT NO. 1
USE AND CARE OF THE MICROSCOPE

Name: ______________________ Rating: __________

Date: ____________

_________________________________________________________________________________

Results and Illustrations:

Draw the microscope and labels it parts. State the function of each part of the microscope. (Use
an extra sheet of bond paper)

Eyepiece Lens
Tube

Coarse Focus

Nosepiece Fine Focus

Rack Stop
Objective Lenses
Arm

Stage

Illuminator Base
Eyepieces: The eyepieces are the lenses at the top that the viewer looks through; they are usually 10X or
15X. To get the total magnification level, multiply the magnification of the objective used (ex: 10X
eyepiece * 40X objective = 400X total magnification).

Tube: Where the eyepieces are dropped in. Also, they connect the eyepieces to the objective lenses.

Base: The bottom of the microscope—what the microscope stands on.

Arm: Structural element that connects the head of the microscope to the base.

Coarse Focus: This is the knob on the side of the microscope that moves the objective lens up and down.
It is used in conjunction with the fine focus

Fine Focus: A knob used to fine-tune the focus of a specimen in conjunction with the coarse focus.

Stage: The flat platform that supports the slides. Stage clips hold the slides in place. If your microscope
has a mechanical stage, the slide is controlled by turning two knobs instead of having to move it
manually. One knob moves the slide left and right, the other moves it forward and backward.

Illuminator: A steady light source (110 volts in the US) that shines up through the slide. Mirrors are
sometimes used in lieu of a built-in light. If your microscope has a mirror, it is used to reflect light from
an external light source up through the bottom of the stage.

Nosepiece: This circular structure is where the different objective lenses are screwed in. To change the
magnification power, simply rotate the turret.

Objective Lenses: Usually you will find 3 or 4 objective lenses on a microscope. The most common ones
are 4X (shortest lens), 10X, 40X and 100X (longest lens). The higher power objectives (starting from
40x) are spring loaded. Spring loaded objective lenses will retract if the objective lens hits a slide,
preventing damage to both the lens and the slide. All quality microscopes have achromatic, parcentered,
parfocal lenses. In addition, to get the greatest clarity at high levels of magnification, you will need a
microscope with an Abbe condenser. Lenses are color coded and are interchangeable between
microscopes if built to DIN standards.

Rack Stop: This feature determines how far up the stage can go. Setting the rack stop is useful in
preventing the slide from coming too far up and hitting the objective lens. Normally, this adjustment is set
at the factory, and changing the rack stop is only necessary if your slides are exceptionally thin and you
are unable to focus the specimen at higher powers.
Draw a few representative cells of the bacteria and show how they appeared at each magnification. Note
the difference in size at each magnification.

Cocci

10X(LPO) 40X/60X(HPO) 100X(OIO)

Bacilli

10X(LPO) 40/60X(HPO) 100X(OIO)

Spirilli

10X(LPO) 40/60X(HPO) 100X(OIO)

REVIEW QUESTIONS?

1. What does it mean when a microscope is parfocal?

It generally means that the objectives can be changed with little to no need of refocusing which
means that if you were to change magnification it will remain focused even when changed

2. Which objective focuses closest to the slide?

100x Oil immersion

3. What controls the amount of light reaching the ocular lens?

The Iris Diaphragm

4. Is your lens corrected for chromatic aberrations? Why or Why not?

Yes, but would highly depend on the on the light that is being used on the microscope, due to the
installable achromatic lenses (blue or red) or that modem that has pre-installed achromatic lenses
already

5. Name two ways in which you can enhance the resolving power?

a. Use an objective lens with the same magnification but larger in diameter.

b. Use Immersion oil________________________________________

6. What advantage does the low power objective have over the oil immersion objective?

Low power is used to see the general picture of the specimen. Also, it is used to find which
portion of the specimen to focus on, after which you change to oil immersion objective

7. What could occur if water were accidentally used in place of immersion oil?

The Water would not aid in the resolving power of the microscope, rather it would blur that part
of the specimen rather than focus it