BIOTECHNOLOGY

Laboratory Activity

Microscope

Group No.: ______

Name: _______________________________________________________________________________

    "Micro" refers to tiny, "scope" refers to view or look at. Microscopes are tools used to enlarge images of small objects
so as they can be studied. The compound light microscope is an instrument containing two lenses, which magnifies, 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 being a light microscope.  In this lab, we will learn about the
proper use and handling of the microscope.

Objectives:

 Demonstrate the proper procedures used in correctly handling using the compound light microscope.
 Prepare and use a wet mount.
 Determine the total magnification of the microscope.
 Describe changes in the field of view and available light when going from low to high power using the
compound light microscope
 Explain why objects must be centered in the field of view before going from low to high power using the compound light
microscope.
 Explain how to increase the amount of light when going from low to high power using the compound light microscope.
 Explain the proper procedure for focusing under low and high power using the compound light microscope.
 Observe onion cells under a microscope.

Materials:

Compound microscope Scissors

Glass slides Iodine solution/povidone iodine
Cover slips Onion bulb
Eye dropper Scalpel/cutter/blade
Beaker of water Forceps
Dissecting needle
The letter "e" cut from newsprint
Tissue paper
Procedure

I. Microscope Parts and Function

1. Carry the microscope with both hands --- one on the arm and the other under the base of the microscope.
2. One person from each group will now go over to the microscope storage area and properly transport one
microscope to your working area.
3. The other person in the group will pick up a pair of scissors, newsprint, a slide, and a cover slip.
4. Remove the dust cover and store it properly. Plug in the scope. Do not turn it on until told to do so.
5. Examine the microscope and give the function of each of the parts listed on the right side of the diagram. Use a
separate sheet to list and define the function of each part of the microscope.

1. eyepiece or ocular
2. body tube
3. fine adjustment knob
4. nosepiece
5. high power objective
6. low power objective
7. diaphragm
8. mirror (many microscopes have a light
instead)
9. base
10. coarse adjustment knob
11. arm
12. stage clip 
13. inclination joint
  

Part II. Preparing a wet mount of the letter "e”.

1. With your scissors cut out the letter "e" from the newspaper.
2. Place it on the glass slide so as to look like (e).
3. Cover it with a clean cover slip. See the figure below.

4. Using your eyedropper, place a drop of water on the edge of the cover slip where it touches the glass slide.
The water should be sucked under the slide if done properly.

Technique for Adding a Stain when making a Wet Mount

5. Turn on the microscope and place the slide on the stage; making sure the "e" is facing the normal reading
position (see the figure above). Using the course focus and low power, move the body tube down until the "e"
can be seen clearly. Draw what you see in the space below.

6. Describe the relationship between what you see through the eyepiece and what you see on the stage.
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7. Looking through the eyepiece, move the slide to the upper right area of the stage. What direction does the image
move?
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8. Now, move it to the lower left side of the stage. What direction does the image move?
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9. Re-center the slide and change the scope to high power. You will notice the "e" is out of focus. DO NOT touch the
coarse focus knob, instead use the fine focus to resolve the picture. Draw the image you see of the letter e (or
 part of it) on high power.

10. Locate the diaphragm under the stage. Move it and record the changes in light intensity as you do so.
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III. Determining Total Magnification:

1. Locate the numbers on the eyepiece and the low power objective and fill in the blanks below.

Eyepiece Objective Total

magnification X magnification = Magnification
______________ ______________ _____________

2. Do the same for the high power objective.

Eyepiece Objective Total

magnification X magnification = Magnification
______________ ______________ _____________

3. Write out the rule for determining total magnification of a compound microscope:

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4. Remove the slide and clean it up.

IV. Examining the Onion Tissue

1. Peel the outer skin of the onion bulb and discard.

2. Using a cutter/scalpel, cut the first layer of the onion bulb and peel. Discard the first layer.
3. Cut a small part of the second layer. Check for the thin layer below.
4. Peel a translucent piece of tissue underneath the second layer of the onion. (The smaller the piece the better.)
Translucent means that you can see light through the specimen, but it is not transparent.
5. Place the piece of onion on a glass slide and add a drop or two of the iodine solution. Cover the slide with a cover
slip using your best wet-mount making techniques.
6. Observe the onion cell under both low and high power. Make a drawing of one onion cell, labeling all of its parts
as you observe them. (At minimum you should observe the nucleus, cell wall, and cytoplasm.)
7. Draw and label the onion cell at both low and high power:
8. Remove the slide and clean it up. Turn off the microscope and wind up the wire so it resembles its original
position. Place the low power objective in place and lower the body tube. Cover the scope with the dust cover. Place
the scope back in its original space in the cabinet

Conclusion Questions:

1. State TWO procedures that should be used to properly handle a light microscope.
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2. Explain why the light microscope is also called the compound microscope.
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3. Images observed under the light microscope are reversed and inverted. Explain what this means.
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4. Explain why the specimen must be centered in the field of view on low power before going to high power.
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5. A microscope has a 20 X ocular (eyepiece) and two objectives of 10 X and 43 X respectively.
a) Calculate the low power magnification of this microscope. Show your formula and all work inside the first
box.
b) Calculate the high power magnification of this microscope. Show your formula and all work inside the second
box

6. In three steps using complete sentences, describe how to make a proper wet mount of the letter e.
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7. Describe the changes in the field of view and the amount of available light when going from low to high power
using the compound microscope.
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8. Explain what the microscope user may have to do to combat the problems incurred in question # 7.
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