Microscope Lab Handout BIO 168

When things are too small to view with just the human eye, a compound light microscope is a tool that
can be used to magnify the specimen so that it can be viewed. A compound light microscope uses two
types of lenses and has its own light source. The intent of this lab is to help you gain an understanding
of basic microscopy.

I. Parts of a microscope
a. Three regions of the microscope
i. Head (or body – this is the top region of the microscope.
ii. Arm – this is the middle region of the microscope
iii. Base – this is the bottom region of the microscope

b. Ocular lens – this is the lens also known as the “eyepiece”. This is what you look
through. On the scopes used in most college classrooms, they have a magnifying power
of 10x.

c. Objective lens – this lens is close to the object which is being magnified. Most
microscopes will have three or four objective lenses that can be rotated.
i. Scanning objective – this lens has a magnifying power of 4x.
ii. Low power objective – this lens has a magnifying power of 10x.
iii. High power objective – this lens has a magnifying power of 40x.
iv. Oil immersion objective – this lens has a magnifying power of 100x but requires
oil for its proper use.

d. Revolving nosepiece – this structure houses the objective lenses. It rotates so that the
objective lenses can be easily changed in order to increase or decrease magnification.

e. Stage – platform on which the specimen is placed

i. Mechanical stage – Most microscopes have stages known as “mechanical
stages”. The mechanical stage allows for the specimen to be moved more easily.
It allows for the specimen to be moved left/right and forward/backward by
turning knobs known as “stage knobs”.

f. Stage knobs – These two knobs allow for the stage to move left/right and
backward/forward.

g. Coarse focus knob – This knob moves the stage up/down. It is used for the initial
focusing of a specimen on the lowest magnification. It should only be used when the
scanning objective lens is in place.

h. Fine Focus knob – This know moves the stage up/down for fine focusing after the
specimen has been brought into focus using the coarse focus knob. It should be the
only focus knob used on low power, high power, and oil immersion settings.

i. Condenser – collects and focuses the light shining up through the specimen

j. Iris diaphragm lever – located on the condenser and controls the amount of light
passing through the specimen

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 Microscope Lab Handout BIO 168

https://upload.wikimedia.org/wikipedia/commons/4/42/OSC_Microbio_02_03_Brightfiel.jpg

II. How to compute total magnification

Both the ocular lens and the objective lens have magnifying power. They work together to
produce total magnification. The formula for determining total magnification is:

Total magnification = ocular lens magnification multiplied by objective lens magnification

III. What is the field of view?

When you are looking through the ocular lens, what you see is called the “field of view”. As
you increase magnification, your field of view gets smaller. So, when using the scanning
objective lens, the field of view is largest, and you have the best chance of locating
specimen. When the specimen is viewed in the field of view, it is upside down and
backwards. If you move the stage to the left, it will appear to move to the right in the field
of view. If you move the stage forward, it will appear to move backward in the field of view.

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IV. How to focus a microscope

a. Always start on scanning objective lens.
b. Use coarse focus knob on scanning objective lens. Do NOT use coarse focus on low
power, or high power, or on oil immersion lenses.
c. Center object before moving up in magnification.
d. Use only fine focus on low power and high power.

V. What does parfocal mean?

Parfocal means that when a specimen is in focus under one objective lens, then it will be
close to being in focus under the other objective lenses. This is why it is only necessary to
use the fine focus knob on higher magnifications (after the specimen is focused under the
scanning objective lens).

VI. How to store a microscope.

a. Lowest power objective lens in place
b. Stage completely lowered
c. Light source turned completely down and off

VII. Working distance.

The working distance is the distance between the top of the slide sitting on the stage and
the bottom of the objective lens. When storing a microscope, you want to have the largest
possible working distance.

VIII. Videos to watch to facilitate understanding of the lab material

a. https://youtu.be/ZblniLyI8uo Parts of the light microscope
b. https://youtu.be/3YgaCH60rQ8 Use and care of the microscope
c. https://yutu.be/vKnypybq7L0 Viewing the letter “e”

IX. Complete the virtual lab on McGraw-Hill Connect.

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 Microscope Lab Handout BIO 168

Lab Assessment 2
1. Complete the following table
Objective Lens in Ocular Lens Objective Lens Total Magnification
place Magnification Magnification

Scanning
Low-power
High-power
Oil-immersion

2. Which objective lens has the largest field of view? ____________________________________

3. Which objective lens has the smallest field of view? ___________________________________

4. If you viewed a piece of paper with the word “eye” using a microscope, how would it look in the
field of view? Draw your answer in the circle below.

5. Which objective lens has the largest working distance? ________________________________

6. Which objective lens has the smallest working distance? ________________________________

7. When viewing a specimen using the low-power objective lens, which focus knob should you be
using? ________________________________________________
8. Why should you center an object before you move up in magnification?
____________________________________________________
____________________________________________________
9. Complete the virtual lab on McGraw-Hill Connect titled “Microscopy – Operation of Brightfield
Microscope”.