Name: _______________________________​ ​ ​ ​ Date: _____________

Introduction to Microscopes Lab

(Type your answers in Blue Font)
Introduction
"Micro" means tiny, and "scope" means to view or look at. Microscopes are tools used to enlarge images of
small objects so they can be studied. The compound light microscope is an instrument containing two lenses,
which magnify, and a variety of knobs to resolve (focus) the picture. Because it uses more than one lens, it is
sometimes called the compound microscope, in addition to being referred to as a light microscope. In this lab,
we will learn how to properly use the compound light microscope.

Goals of this Lab:

●​ Identify all of the parts of a compound light microscope.
●​ Describe the function of each of the parts of the compound light microscope.
●​ Demonstrate the proper procedures in correctly using the compound light microscope.
●​ Determine the total magnification for each objective lens on the microscope.
●​ Describe changes in the field of view and available light when going from low to a higher power using
the compound light microscope.
●​ Explain why objects must be centered in the field of view before going from low to a higher power
using the compound light microscope.
●​ Explain how to increase the amount of light when going from low to higher power using the compound
light microscope.
●​ Demonstrate the proper procedure for focusing under low power, medium power, and high power using
the compound light microscope.
●​ Prepare and view a wet mount on a slide.

Part I: Microscope Handling, Parts, and Functions

1.​ Either you or your partner need to obtain a microscope from the storage closet. Carry the microscope
with both hands - one hand on the arm and the other under the base of the microscope.
2.​ Remove the dust cover and store it properly. Plug in the scope. Do not turn it on until told to do so.
3.​ Examine the microscope and identify each of the parts and its function with your lab partner.

Part II: Determining Total Magnification

The microscope is a device used to make very tiny (microscopic) objects visible to the human eye through
magnification. The very first microscopes had only one lens, and were referred to as simple microscopes.
Compound microscopes have at least two lenses and were invented in the 1590s. The first microscopes relied on
light to see the sample being viewed and were called optical or light microscopes.

Compound light microscopes like we use in Biology rely on light to see the sample and have more than one lens
to magnify the sample. Typically, compound light microscopes have an ocular lens, also called the eyepiece,
and 3-4 objective lenses. The ocular lens on our microscopes has a magnification of 10X. Look on the ocular
lens and locate the magnification. Write this ocular lens magnification in the chart below.
Now look at each of the objective lenses and locate their magnification. Fill in the magnification for each
objective lens in the chart below. To calculate total magnification on a microscope, you must consider both the
magnification of the ocular lens and magnification of the objective lens that is placed over the sample. The
equation to determine total magnification is: ​

Magnification of Ocular Lens X Magnification of Objective Lens = Total Magnification. ​

Use this equation to answer questions 1 and 2 below.

Ocular/Eyepiece Objective Lens Total Magnification

Power Objective
Magnification Magnification (show your calculation)

Low (Scanning) 10x 4x 40x

Medium 10x 10x 100x

High 10x 40x 400x

Oil Immersion

Answer the following questions:

1.​ What is the equation for determining total magnification for a compound light microscope?
​
Magnification of the eyepiece lens X Magnification of the objective lens = Total Magnification.

2.​ Use the equation from question (1) to calculate the total magnification for each of the objective lenses in
the chart above. Record the total magnification in the chart.

Part III: Preparing a wet mount of the letter "e”

1.​ With your scissors, cut out a typical sized letter "e" from the newspaper. Leave a couple of letters before
and after and above and below the “e” so that it is easier to manipulate with your forceps.
2.​ Pick up the “e” with your forceps and place the letter “e” on the glass slide right side up as if you were
reading it (e).
3.​ Use the dropper to place one drop of water on the “e”. Use forceps to make sure that the “e” is still
positioned as if you are reading it.
4.​ Cover the “e” with a clean cover slip. See the figure below. ​

5.​ Move your stage all the way down and rotate the objective lenses so that the low/scanning objective lens
is above the hole on your stage.
6.​ Pull back the stage clip and place the slide on the stage making sure the "e" is facing the normal reading
position (see the figure above) and is above the hole on the stage.
a.​ Do NOT put the slide under the stage clips. This will break the microscope.
b.​ The diagram below shows what your slide should look like when loaded.​

7.​ Turn on the microscope.

8.​ Using the coarse focus adjustment knob, move the stage up until the "e" can be seen clearly through the
eyepiece. Draw what you see below. Include the total magnification at which you view your specimen.​
​
Draw the letter “e” as seen through your microscope on low power.
​

​ ​ Total Magnification: 40x​

9.​ How does the way you see the “e” through the microscope differ from the way you see it without the
microscope? Do they look the same? Different? How so?

The “e” was upside down and backwards when we viewed it through the microscope. ​

10.​Look through the eyepiece and move the slide toward you using the stage dials. What direction does the
image move? ​
 When sliding it towards me, the letter “e” moves in the opposite direction, away from me. ​

11.​Now, move the slide to the left. What direction does the image move?
​
The image moves to the right when you move the slide leftways. ​

12.​Move the stage to the right. What direction does the image move?

The image moves to the left when you move the slide to the right. ​

13.​Re-center the slide so your “e” is in the center of your field of view. Refocus using your coarse
adjustment knob and sharpen the focus using your fine focus adjustment knob.
14.​Change the scope to medium power. You will notice the "e" is out of focus. Do not touch the coarse
focus adjustment knob. Instead use the fine focus adjustment knob to sharpen your focus. Draw the
image you see of the letter “e” (or part of it) on medium power below. Include the total magnification at
which you view your specimen. Be sure your drawing represents what the letter “e” actually looks like
when magnified. ​
​
Draw the letter “e” as seen through your microscope on medium power.
​

​ ​ Total Magnification:100x​

15.​Locate the diaphragm under the stage. Move it. What changes do you observe in light intensity as you
move the diaphragm?

Moving the diaphragm to the right decreases the light intensity whereas moving it to the left will
increase the light intensity slightly. ​

16.​Turn off the microscope. Change the objective lens to low power. Use the coarse adjustment knob to
lower the stage completely. Pull back the stage clip and remove your slide.
Part IV: Looking at Prepared Slides
1.​ Choose one of the prepared slides. Write what slide you are viewing below. ​
Slide: 91 W 7940 - Ligustrum Leaf ​

2.​ Using low power, draw what you see. Include the total magnification.
​

​ ​ Total Magnification: 40x​

​

3.​ Repeat looking at the slide with medium power. Remember to focus your specimen in the center of your
field of view in low power first, then change to medium power. Sharpen the focus of your image using
the fine focus adjustment knob only. Use your diaphragm to adjust the amount of light shining through
the sample to get the best image possible. Draw what you see and include the total magnification below.

​ ​ Total Magnification: 100x​

4.​ Repeat step 3 by making sure your specimen is in the center of your field of view on medium power and
that it is sharply focused (fine focus adjustment knob only), and move your objective lens to high power.
Sharpen the focus of your image using the fine adjustment knob only. Use your diaphragm to adjust the
amount of light shining through the sample to get the best image possible. Draw what you see and
include the total magnification.
​

​ ​ Total Magnification: 400x​

5.​ Turn off the microscope. Change the objective lens to low power. Lower the stage completely. Pull back
the stage clip and remove the slide. Return your slide to Mr. Nadar
Part V: Clean-Up
Turn off the microscope, unplug and wind up the cord so it resembles its original position. Lower the stage and
place the low/scanning power objective over the hole in the stage. Cover the scope with the dust cover. The
partner that did not get the microscope at the beginning of this lab will return the microscope to the storage
closet, taking care to carry the microscope properly.

Conclusion Questions:
1.​ State two procedures which should be used to properly handle a light microscope. ​
Procedure 1: Make sure that the light is turned off and the stage is lowered fully before placing the cover
back on. ​
​
Procedure 2: When picking up the microscope, make sure to have one hand on the arm and another on
the base for maximum safety. ​

2.​ Images observed under the light microscope are reversed and inverted. Explain what this means. Include
a picture as part of your explanation. ​
​

Picture:
This happens due to the optical part of a microscope, specifically the lens and the way they are arranged.
The objective lens and eyepiece lens create a picture that will be flipped both horizontally and vertically. ​

3.​ Explain why the specimen must be centered in the field of view on low power before moving to medium
power. ​
​
It is important to center the specimen when switching over to a medium power because it has a narrower
field of view, therefore, most of the time, making it easier to lose sight of the specimen. Adjusting the
specimen also helps get a clearer view and lowers the risks of damaging the slide or specimen.

4.​ Suppose a microscope has a 20 X ocular (eyepiece) and low and medium power objective lenses with 10
X and 43 X magnification, respectively: ​
a. Calculate the low power magnification of this microscope. Show your formula and all work. ​

20 (eyepiece) x 10 (low power) = 200x total zoom ​

​
 ​
b. Calculate the medium power magnification of this microscope. Show your formula and all work. ​
​
20 (eyepiece) x 43 (medium power) = 860x total zoom ​
​

5.​ Describe how each of the following change when moving from low to medium power when using the
compound microscope: ​
a. the field of view ​
​
When moving from low to medium power, the field of view will decrease. Therefore, only a smaller
area of the specimen will be focused on, allowing for a more detailed and precise image of the features. ​
​
b. the amount of available light ​
​
When increasing the power, the amount of light able to reach it will decrease, that means that higher
magnifications will need more light to maintain a clear image. Users may need to increase the light
intensity for a clearer view of the specimen. ​

6.​ Explain what you should do to avoid problems that may happen to your field of view and the amount of
light available when moving from low power to medium power. ​
​
To avoid any problems that may happen when switching from low to medium power, users should make
sure that the specimen is still centered and to increase the light intensity. Another thing users should do
is to use the focus knob to allow for more focus on the image without any damage being done to the
slide or lens.

7.​ How does the procedure for focusing the image differ under high power compared with low power?

The main difference between focusing an image under high power and low power is that users will use
the fine focus knob during the high power mode to avoid any damage being done to the microscope lens,
whereas, users would use the coarse focus knob for low power scanning. Another big difference is that
users will highly need to increase the light intensity, while not needing to do the same thing with low
power.