EXERCISE 2: USE AND CARE OF

THE MICROSCOPE

Introduction:
The bright-field light microscope is an instrument that magnifies images using two-lens
system. Initial magnification occurs in the objective lens. Most microscopes have at least
three objective lenses on a rotating base, and each lens may be rotated into alignment with the
eyepiece or ocular lens in which the final magnification occurs. The objective lenses are
identified as the low-power, high-dry, and oil immersion objectives. As the magnification
increases, the size of the lens at the tip of the objective becomes progressively smaller and
admits less light. This is one of the reasons that changes in position of the sub-stage
condenser and iris diaphragm are required when using different objectives if the specimens
viewed are to be seen distinctly. When oil immersion lens is used, immersion oil fills the
space between the objective and the specimen. Because immersion oil has the same
refractive index as glass, the loss of light is minimized.

Purpose:
The purpose of this exercise is to introduce students with the proper care and use of the
compound light microscope.

Objectives:
 Be able to familiarize the parts of the microscope and the function of each part.
 Be able to explain the proper usage and care of the microscope.
 Be able to demonstrate the proper usage of the microscope by locating, focusing, and
identifying a specimen on a microscope slide.
 Be able to determine the total magnification of the specimens under the microscope.

Instructions:

Please go to this website https://virtuallab.nmsu.edu/stain.php where you will be redirected to

the virtual lab simulations. Be sure to click “Using the Microscope” section at the upper part
of the page. Follow the instructions in the virtual lab and learn the basics of microscopy.

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A. Microscope Labeling

Eyepiece/Ocular Lens

Body Tube

Nosepiece
Arm
Low-power
Objectives (10x)
High Dry Objectives
(40x)

Oil immersion Stage

Objectives (100x)

Stage Clips Coarse Adjustment Knob

Diaphragm Fine Adjustment Knob

Light Source
Base

Microscope Use:

15. When focusing a specimen, you should always start with the 10x
objective.
16. When using the high power objective, only the Fine adjustment knob should be
used.
17. Course adjustment knob is used for bringing the specimen into focus, and
Fine adjustment knob is used for sharpening the image of the specimen after it is
focused.
18. You should carry the microscope by the Arm and the Neck.
19. The objectives are attached to what part of the microscope (it can be rotated to click
lenses into place?) Nosepiece
20. What is the function of the diaphragm? It adjusts the amount of light on the slide.
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 B. Calculation of Total Magnification:

Magnification is the number of times an image is enlarged or reduced by a lens system or by

a drawing. Compound microscopes typically have at least 4 lenses. The ocular (eyepiece) lens
usually magnifies the image ten times, or at 10x. The 3 objectives often magnify at 4x, 10x,
and 40x, though each microscope is different. To calculate the magnification, simply multiply
the magnifying power of the ocular lens (10x) with the magnifying power of the objective
lens used. With this microscope you can obtain three different magnifications.

Answer the proceeding questions briefly but concisely.

1. In your own words, how do you calculate total magnification?

You should multiply the occular lens power to the objective lens power. The standard eyepiece magnifies
10x. For example let’s multiply the Oil immersion power which is 100x and with the eyepiece which is 10x
That is equal to 1000x.

2. Calculate the total magnifications for scanning, low, high and OIO views:

Common
Total Magnification
Objective Power Magnifications of
(Ocular Lens = 10x)
these Lenses
Scanning 4X 40x
Low 10X 100x
High 40X – 45X 400x-450x
Oil Immersion 100X 1000x

C. Specimen observations: Go to section D and examine all the specimen images. Draw
only two settings per specimen. Be sure to label your images with the magnification and
the name of the microorganism.

Common Name: Alga Common Name: Alga

Scientific Name: Spirogyra Scientific Name: Spirogyra
Magnification: 100 X Magnification: 400 X
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Common Name: Bacterium Common Name: Bacterium
Scientific Name: Lactobacillus Scientific Name: Lactobacillus
Magnification: 40 X Magnification: 400 X

Common Name: Protozoan Common Name: Protozoan

Scientific Name: Amoeba Scientific Name: Amoeba
Magnification: 40 X Magnification: 400 X

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D. Images of specimens examined under the different objective lenses of a compound light
microscope.

Amoeba species under LPO Amoeba species under MPO

Amoeba species under HPO

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Lactobacillus species under LPO Lactobacillus species under MPO

Lactobacillus species under HPO

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 Spirogyra species under MPO Spirogyra species under HPO

REFLECTION/GENERALIZATION:

It is encouraged and vital to use a microscope when studying microorganisms because there will be very little to do
in the laboratory without it. The microorganism would be so small to see that it would most likely be unrecognizable.
Given the tiny size of these creatures, it is preferable and more accurate to view as well as their shape and color and
it is just possible when a microscope is used. We are able to see in detail what our eyes could not perceive. We've also
learned how important it is to properly maintain and use the microscope, one of which is to carry it properly by
holding it with both hands and assisting the other hand on the base of the microscope while the other one carries the
arm.

References:
 Compound Microscope Parts. (2021). Microscope.Com.
https://www.microscope.com/compound-microscope-parts
 Amoeba | Encyclopedia.com. (2021). Encyclopedia.Com.
https://www.encyclopedia.com/plants-and-animals/microbes-algae-and-fungi/
moneran-and-protistan/amoeba
 Spirogyra | Definition, Structure, Reproduction, & Facts. (2021). Encyclopedia
Britannica. https://www.britannica.com/science/Spirogyra

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