Name: _______________________________Date:____________ Score: ______________

Group Number: ____ Section: __________ Instructor’s Signature: ____________________

Laboratory Activity No. 1

Microscopy

Introduction

Microscopy is the use of microscopes to study objects that are too small to see with
the naked eye. It's a key tool in microbiology, which is the study of the structure and function
of microscopic living things.
Microscopy is used by scientists and health care professionals for many purposes,
including diagnosis of infectious diseases, identification of microorganisms (microscopic
organisms) in environmental samples (including food and water), and determination of the
effect of pathogenic (disease-causing) microbes on human cells. This exercise will
familiarize you with the microscopes we will be using to look at various types of
microorganisms throughout the semester.

Objectives

1. Demonstrate proper use, care and transport of the microscope.

2. Determine the total magnification of the microscope.
3. Explain why objects must be centered in the field of view before going from low to high
power using the compound light microscope.
4. Explain how to increase the amount of light from low to high power using the compound light
microscope.
5. Using the compound light microscope, explain the proper procedure for focusing under low
and high power.

Materials & Apparatus

Compound microscope, glass slides, cover slips, forceps
Sample specimen: hair, leaf, fiber/ thread, water, soil, onion skin,Letter ‘e’ cut out

Procedure.

1. Familiarize the parts of the microscope and its function. Review the parts of the microscope,
making sure you know their names and understand the function of each of these components.
2. Review instructions for the use of the microscope,giving special attention to the use of the
low power , high power and oil-immersion objectives.
3. Examine the prepared slides, noting the shapes and the relative sizes of the cells under
the
high-power (also called high-dry, because it is the highest power that does not use oil) and
oil immersion objectives.
4. Record your observations in the Lab Report.

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Part I. A. Parts and Function of a Microscope

The Compound Microscope and Its Parts

B. Use and Care of the Microscope.

B.1. Carrying the Microscope from one place to another.

1. Often, you must move microscopes to your laboratory bench/table. The correct way to do
this is to grip the microscope arm firmly with one hand and the base with your other hand to
lift the instrument.
2. Carry it close to the body and gently place it on the laboratory bench. This will prevent
collision with furniture or coworkers and will protect the instrument against damage.

B.2. Cleaning the Microscope and its lenses.

Once the microscope is placed on the laboratory bench, observe the following rules:
1. Remove all unnecessary materials (including books, papers, purses,bags and hats) from
the laboratory bench.
2. Uncoil the microscope’s electric cord and plug it into an electrical outlet.
3. Clean all lens systems; the smallest bit of dust,oil, lint, or eyelash will decrease the
efficiency of the microscope. The ocular, scanning, low power,and high-power lenses may
be cleaned by wiping several times with acceptable lens tissue. Never use a paper towel or
cloth on a lens surface. If the oil-immersion lens is gummy or tacky, ask your instructor to
use a piece of lens paper moistened with xylene to wipe it clean. Your instructor should

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immediately remove the xylene with a tissue moistened with 95% alcohol, and wipe the lens
dry with lens paper.

B.3. Focusing and Viewing Specimens on the Microscope.

1. Place the microscope slide with the specimen within the stage clips on the fixed stage.
Move
the slide to center the specimen over the opening in the stage directly over the light source.
2. Raise the microscope stage up as far as it will go.
3. Rotate the scanning lens or low-power lens into position.
4. Lower the body tube with the coarse-adjustment knob to its lowest position.
Note: Never lower the body tube while looking through the ocular lens; this may allow for an
impact with the slide and damage to the slide or the microscope.
5. While looking through the ocular lens, use the fine-adjustment knob, rotating it back and
forth slightly, to bring the specimen into sharp focus.
4. Adjust the substage condenser to achieve optimal focus.
5. Routinely adjust the light source by means of the light-source transformer setting, and/or
the iris diaphragm, for optimum illumination for each new slide and for each change in
magnification.
6. Most microscopes are parfocal, which means that when one lens is in focus, other lenses
will also have the same focal length and can be rotated into position without further major
adjustment. In practice, however, usually a half-turn of the fine-adjustment knob in one
direction or the other is necessary for sharp focus.
7. Once you have brought the specimen into sharp focus with a low-powered lens, prepare
to visualize the specimen under high power by rotating the high power lens into
position.Readjust the fine-adjustment knob to bring the image into sharp focus.
8. When using the oil immersion lens (100X), place a drop of oil on the slide directly over
the viewing area.
9. Rotate the nose piece until the oil-immersion objective locks into position.
Note: Care should be taken not to allow the high-power objective to touch the drop of oil.
Observe the slide from the side as the objective is rotated slowly into position. This will
ensure that the objective will be properly immersed in the oil. Readjust the fine-adjustment
knob to bring the image into sharp focus.
10. During examination of microbial organisms, it is always necessary to observe several
areas of the preparation. To do so,scan the slide without the application of additional
immersion oil. Note: This will require continuous, very fine adjustments by the slow, back-
and-forth rotation of the fine adjustment knob only.

B.4 After use care of the Microscope.

On completion of the laboratory exercise,return the microscope to its original location and
condition. The following steps are recommended:
1. Clean all lenses with dry, clean lens paper.
Note: Use xylene to remove oil from the stage only.
2. Place the low-power objective in position and lower the body tube completely.
3. Center the mechanical stage.
4. Coil the electric cord around the body tube and the stage.
5. Carry the microscope to its position in its cabinet carefully, as previously described.

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 B. Magnifying Power of the Microscope
Magnification is the ratio of the image size with the microscope to the actual size of
the object. When you say that the total magnification is 10x, the image you see using the
microscope is ten times bigger than viewing the specimen with the naked eye. Remember
with a compound light microscope you are magnifying with two lenses, so to calculate
the total magnification, you multiply the ocular or eyepiece lens magnification by the
objective lens magnification.
For example, if the objective lens is 40x and the eyepiece lens is 10x,
40x multiplied by 10x = 400x is the total magnification.

Complete the table below:

Part II.
A. Procedure for 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 to look like (e).
3. Cover it with a clean cover slip.
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.
5. Turn on the microscope and place the slide on the stage; making sure the "e" is facing the
normal reading position. Using the coarse focus and low power, move the body tube down until
the "e" can be seen clearly.
6. Draw what you see in the space below in your lab report.
7. Looking through the eyepiece, move the slide to the upper right area of the stage. What
direction does the image move? _____________________
8. Now, move it to the lower left side of the stage. What direction does the image move?
________________.
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 in your lab report.
10. Locate the diaphragm under the stage. Move it and record the changes in light intensity as
you do so.
B. Observe the following specimen under the microscope:
a. onion skin
b. Thread
c. All specimen on hand-hair, fiber, water, soil,Letter ‘e’ cut out

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 Name: _______________________________Date:____________ Score: ______________
Group Number: ____ Section: __________ Instructor’s Signature: ____________________

Laboratory Activity No. 1

LABORATORY REPORT
Microscoppy
Questions:
1. Draw a microscope and label the parts.State the function of each part.Use the back portion of
this page for the functions.

2. State 2 procedures that should be used to carry/handle a light microscope properly.

______________________________________________________________________________
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3. Explain why the light microscope is also called the compound microscope.
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4. Draw the letter ‘e’ as viewed on low power objective and high power objective.

Low power High Power

Magnification: Magnification

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5. Images observed under the light microscope are reversed and inverted. Explain why.
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6. Explain why the specimen must be centered in the field of view on low power before going to
high power.
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7. 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 your
solution.
______________________________________________________________________________
______________________________________________________________________________

b) Calculate the high power magnification of this microscope. Show your formula and your
solution.
______________________________________________________________________________
______________________________________________________________________________

8.In three steps using complete sentences, describe how to make a proper wet mount of the letter
“e”.
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
9. How does the procedure for using the microscope differ under high power as opposed to low
power?
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______________________________________________________________________________
______________________________________________________________________________

CONCLUSION. (Based your discussion from the objectives of this experiment)

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Forensic Chemistry College of Criminology St. Michael’s College Expt. 1 6