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Proper Use and Handling of Microscope

I. Introduction:
The applications of microscopy in the forensic sciences are almost limitless. This is due in large
measure to the ability of microscopes to detect, resolve and image the smallest items of evidence, often without
alteration or destruction. As a result, microscopes have become nearly indispensable in all forensic disciplines
involving the natural sciences. Thus, a firearms examiner comparing a bullet, a trace evidence specialist identifying
and comparing fibers, hairs, soils or dust, a document examiner studying ink line crossings or paper fibers, and a
serologist scrutinizing a bloodstain, all rely on microscopes, in spite of the fact that each may use them in different
ways and for different purposes.

II. Objectives:

After the activity, you should be able to:

1. Identify the parts of the microscope and their functions.
2. Become familiar with the 3 variations of light microscopy.
3. Determine total magnification of the specimen, using various objective lenses.
4. Be able to switch objective lenses, while focused on the specimen, without moving the stage.
5. Handle the microscope safely and clean it.
6. Explain principles and terms used in microscopy and focusing.
III. Materials:
Microscope glass slide glass cover

IV. Procedure:
A. Safe Handling and Operation
1. When moving your microscope, always carry it with both hands (Figure 1). Grasp the arm with one hand
and place the other hand under the base for support.
2. Keep one hand on the arm, and one on the base. Do not carry anything else with a microscope.
3. Clean ALL objective lenses and the ocular with lens paper BEFORE you even place a slide on the stage, and it
is a good idea to wipe the condenser lens also. The last person using the microscope may have left it dirty:
it is imperative to begin with cleanliness.
4. Turn the revolving nosepiece so that the lowest power objective lens is "clicked" into position.
5. Place the microscope slide on the stage and fasten it with the stage clips. You can push down on the back
end of the stage clip to open it.
6. Using the coarse adjustment, lower the objective lens down as far as it will go without touching the slide!
Note: Look at the slide and lens from the side when doing this (see Figure 2).
7. Look through the eyepiece and adjust the illuminator (or mirror) and diaphragm (Figure 3) for the greatest
amount of light.

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8. Slowly turn the coarse adjustment so that the objective lens goes up (away from the slide). Continue until the
image comes into focus. Use the fine adjustment, if available, for fine focusing.

9. Move the microscope slide around so that the image is in the center of the field of view and readjust the
mirror, illuminator or diaphragm for the clearest image.

10. You should be able to change to the next objective lenses with only slight focusing adjustment. Use the fine
adjustment, if available. If you cannot focus on your specimen, repeat steps 4 through 7 with the higher power
objective lens in place. DO NOT ALLOW THE LENS TO TOUCH THE SLIDE!

11. The proper way to use a monocular microscope is to look through the eyepiece with one eye and keep the
other eye open (this helps avoid eye strain). If you have to close one eye when looking into the microscope, it's
ok. Remember, everything is upside down and backwards. When you move the slide to the right, the image goes
to the left. NOTE: Most microscopes are “parfocal”; that is, the slide should be in focus (or nearly so) when you
switch from low-power to high-power. If you have properly focused your microscope at low-power and your slide
is properly prepared (i.e. the object is flattened by a coverslip), you should be able to switch automatically from
low-power to high without fear of having the high-power objective lens scraping or touching the slide. An
important factor in using the microscope is the distance between the object viewed and the objective lens of the
microscope. This is called the working distance. As you increase magnification the working distance becomes
less.to the left!

12. Do not touch the glass part of the lenses with your fingers. Use only special lens paper to clean the
lenses. (read the page on keeping your microscope clean).

13. When finished, raise the tube, click the low power lens into position and remove the slide.

Remember, microscopes are expensive scientific instruments. Handle them properly and carefully and
they will last for many years!

B. Parts of A Compound Microscope:

1. OCULAR LENS or EYEPIECE —On a binocular scope there are two ocular lenses, one for each eye.
These lenses magnify the image at 10Xpower. The power of the ocular lens multiplied by the
objective lens gives the total magnification of the microscope.
2. ARM —A support for the upper portion of the scope. It also serves as a convenient carrying handle.
3. MECHANICAL STAGE CONTROLS —A geared device to move the slide (placed in the slide clamp)
precisely.
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4. COARSE ADJUSTMENT KNOB —A rapid control which allows for quick focusing by moving the
objective lens or stage up and down. It is used for initial focusing.
5. FINE ADJUSTMENT KNOB —A slow but precise control used to fine focus the image when viewing
at the higher magnifications.
6. BASE —The part of your microscope that sits on a level, stable support.
7. NOSEPIECE —A circular plate with 4 objective lenses that can be rotated into position for different
magnifications.
8. OBJECTIVE LENS —Four separate lenses that magnify the image (4X, 10X, 40X and 100X)
depending on the objective in use. The lens is positioned just above the object being viewed.
OBJECTIVE POWER OBJECTIVE NAME
4X SCANNING
10X LOWPOWER
40X HIGH POWER
100X OIL IMMERSION
9. SLIDE CLIP —A clip to hold the slide on the stage.
10. STAGE —A platform for placement of the microscope slide.
11. CONDENSER —A lens that concentrates or directs the light onto the slide.
12. IRIS DIAPHRAGM CONTROL —A lever (or rotating disk) that adjusts the amount of light
illuminating the slide. Use just enough light to illuminate the object on the slide and give good contrast.
13. LAMP — The light source.
14. LAMP SWITCH —Turns the lamp “on” and “off”.

Draw the Different Parts of A MICROSCOPE

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Guide Questions:
1. Why move the 10X objective lens into place when putting the microscope back into the cabinet?
2 How is the image of an object seen through the high- power objective different from the image seen through
the low-power objective?
3. What would be the total magnification of a specimen using the 40X objective lens?
4. What is parfocal?
5. Why reduce your light when using the 10X objective lens?
6. What happens to the light intensity as you increase magnification?
7. What term is used to describe the feature of the microscope that makes it possible to move among the
objective lenses with just MINOR focusing?
8. Which objective lens is also called the oil-immersion lens?
9. Why do we have to clean all objective lenses before using the microscope?
10.Why do we have to look at the side and not at the center when when lowering the objective lense using the
coarse adjustment knob?

Conclusion: