THE MICROSCOPE

MICROSCOPE BASICS

The microscope must always be handled properly. You must observe the following rules for its
transport, cleaning, use, and storage:
A. Transport in an upright position with one hand on the arm and the other supporting the base.
Set it down carefully at your work station. Do not drag it across the table.
B. Use only special lens paper to clean the lenses. Clean all lenses before and after use.
Slides should also be cleaned.
C. Always begin the focusing process with the 4x or 10x objective lens in position, changing to
the higher-power lenses as necessary.
D. The coarse adjustment knob may be used with the 4x or 10x lens, but use only the fine
adjustment with 40x or 100x.
E. Adjust lighting appropriately. Turn off the light when not in use.
F. Always use a cover slip with temporary (wet mount) preparations.
G. When you put the microscope away, remove the slide from the stage, and rotate the lowest-
power objective lens into position. Wrap the cord around the clips on the back, not around
the base.
H. Never remove or loosen any parts from the microscope.
I. Inform your instructor of any mechanical problems.

ACTIVITY 1: Identifying the Parts of a Microscope

1. Obtain a microscope and bring it to the laboratory bench. (Use the proper transport
technique!) Compare your microscope with the figure on the following page and identify the
following microscope parts:

Base: Supports the microscope. Substage light: Located in the base, the light
passes directly upward through the microscope.

Stage: The platform the slide rests on while being viewed. The stage has a hole in it to
permit light to pass through both it and the specimen. The mechanical stage permits
precise movement of the specimen.

Condenser: Concentrates the light on the specimen. The condenser has a height
adjustment knob that raises and lowers the condenser to vary light delivery. Generally,
the best position for the condenser is close to the inferior surface of the stage.

Iris diaphragm dial: Dial attached to the condenser that regulates the amount of light
passing through the condenser. The iris diaphragm permits the best possible contrast
when viewing the specimen.

Coarse adjustment knob: Used to focus on the specimen when on 4x or 10x.

MC 1L 22/23 1
 Figure 1. The Compound Microscope

Fine adjustment knob: Used for precise focusing once coarse focusing has been
completed. Use only this knob when on 40x or 100x.

Head or body tube: Supports the objective lens system, and the ocular lenses.

Arm: Vertical portion of the microscope connecting the base and the head.

Ocular (or eyepiece): There are two lenses at the superior end of the head, through
which observations are made. An ocular lens has a magnification of 10x. If your
microscope has a pointer, it is attached to the right ocular and can be positioned by
rotating the ocular lens

Nose piece: Has four objective lenses and permits sequential positioning of these
lenses over the light beam passing through the hole in the stage. Use the nose piece to
change the objective lenses.

Objective lenses: Adjustable lens system that permits the use of a scanning lens, a low-
power lens, a high-power lens, or an oil immersion lens. The objective lenses have
different magnifying and resolving powers.
MC 1L 22/23 2
2. Look at the objective lenses carefully. The shortest lens is the scanning lens, and has
magnification of 4x. The low power lens is 10x. The high-power objective lens is 40x. The oil
immersion objective lens is usually the longest of the objective lenses and has a magnifying
power of 100x. Record the magnification of each objective lens of your microscope in the
first row of the summary chart.
3. Rotate the lowest power objective lens until it clicks into position, and turn the coarse
adjustment knob about 180 degrees. Notice how far the stage (or objective lens) travels
during this adjustment. Move the fine adjustment knob 180 degrees, noting again the
distance that the stage (or objective lens) moves.

Table 1. Summary Chart

Scanning Low Power High Power Oil Immersion

Magnification of
Objective lens
Total Magnification

Working Distance

Details observed

MAGNIFICATION & RESOLUTION

The microscope is designed to magnify specimens. Your microscope is called a compound

microscope because it uses two lenses to magnify the specimen. The objective lens magnifies
the specimen to produce a real image that is projected to the ocular. This real image is
magnified by the ocular lens to produce the virtual image seen by your eye.

The total magnification of any specimen being viewed is equal to the power of the ocular lens
multiplied by the power of the objective lens. If the ocular lens magnifies 10x and the objective
lens magnifies 50x, the total magnification is 500x (10 x 50). Determine the total magnification
with each of the objectives on your microscope and record in the chart.

With a compound light microscope such as the one you are using, the level of magnification is
almost limitless, but the resolution (resolving power) is not. Resolution refers to the ability to
discriminate two close objects as separate. The human eye can resolve objects about 100 µm
apart, but the compound microscope has a resolution of 0.2 µm under ideal conditions. Objects
closer than 0.2 µm are seen as a single fused image. Resolving power is determined by the
amount and physical properties of the visible light that enters the microscope.

MC 1L 22/23 3
In general, the more light delivered to the objective lens, the greater the resolution. The size of
the objective lens aperture (opening) decreases with increasing magnification, allowing less light
to enter the objective. You will likely need to increase the light intensity at the higher
magnifications.

ACTIVITY 2: Viewing Objects Through the Microscope

1. Obtain a slide sample of a tissue. Adjust the condenser to its highest position and switch on
the light source of your microscope.
2. Place the slide on the stage (in the slide holder) centered over the light beam passing
through the stage.
3. With your lowest power objective lens in position over the stage, use the coarse adjustment
knob to bring the objective lens and stage as close together as possible.
4. Look through the ocular lens and adjust the light for comfort using the iris diaphragm. Now
use the coarse adjustment knob to focus slowly away from the e until it is as clearly focused
as possible. Complete the focusing with the fine adjustment knob.
5. Sketch what you have observed as it appears in the field (the area you see through the
microscope).
6. Most laboratory microscopes are parfocal. This means the slide should be in focus (or
nearly so) at the higher magnifications once you have properly focused. Without touching
the focusing knobs, increase the magnification by rotating the next higher magnification lens
into position over the stage. Make sure it clicks into position. Using the fine adjustment only,
sharpen the focus. Notice the decrease in working distance. On high power or oil immersion,
focusing with the coarse adjustment knob could drive the objective lens through the slide,
breaking the slide and possibly damaging the lens. What new details can you observe?

Record the detail observed, and working distance in the summary

chart. Guide questions for observation:

A. Is the image larger or smaller than on the previous magnification?
B. Approximately how much of the image is visible now? Is the field larger or smaller?
C. Why is it necessary to center your object (or the portion of the slide you wish to view)
before changing to a higher power?
D. Move the iris diaphragm lever while observing the field. What happens? Is it more
desirable to increase or decrease the light when changing to a higher magnification?
Why?

7. Repeat the steps given in direction #6 using the high-power objective lens. Record the
detail observed, and working distance in the summary chart.
8. We will not use the oil immersion lens today, but you will learn to use it in the next
semester. Never click an oil immersion lens into place without using oil properly. You
should fill in as much of the summary chart as you can right now. What do you think the
working distance should be on oil immersion? What do you think you would observe?
9. Now its time for you to identify the detailed structures of the cell from different tissue
samples. Observe the distinguishing characteristics between cells.
MC 1L 22/23 4