Name: ___________________________________________________ Perfect Rating [At least 107 points]

Yr/Section: ______________________________________________ Date Performed: ______________________

Date Submitted: ______________________

Lab Activity No. 1

Microscopy
Introduction
Cells are too minute to be seen with the naked eye. An instrument like the microscope which can magnify these
cells in a living thing is fundamentally important when studying these structures. "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
 Familiarize yourselves with the microscope.
 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.
 To calculate the size of specimens using calibration factor

Materials
 E-media sources (on microscopy) and other  Red and white threads (2 cm long)
reference materials  Clean Soft cloth
 Microscope  Tissue paper
 Glass slide  Hand towel
 Cover slip
 Medicine Dropper

A. Microscope Handling
1. When carry the microscope with both hands --- one hand should be on the arm and the other under the
base of the microscope.
2. Examine the microscope below. Label the parts and give their functions. (at least 26 points)
B. HANDLING A MICROCOPE
1. Whenever possible, observe proper handling of the microscope.
2. Be acquainted with your microscope’s parts and functions. When using the microscope:
3. Observe the ocular. Look for its magnification etched or printed on its edge. Peer onto the ocular (Make sure to
check your light source if it is turned ON). What you see is your Field of View (FOV). Adjust the light intensity if it
is too bright.
4. Observe and get acquainted with the different kinds of objectives. Locate the scanner, low power objective and
high power objective lens. Check for their individual magnifications etched on their body followed by letter “x”. In
terms of size, scanner is short, LPO is medium and HPO is long. The objectives are held by the revolving nose
piece. Rotate the revolving nosepiece when switching from one objective to other.
5. When mount your specimen onto your slide, cover it with cover slip.
6. Then mount your slide specimen on the microscope stage and secure it using the stage clips.
7. Observe the slide: Use the scanner first. Put the scanner in place. Turn then the coarse adjustment knob clockwise
and counterclockwise. Observe how the objectives are brought towards and away from the slide. Always check
the distance between your slide and the objective lens to prevent breakage of the slide or damage to the objective
lens. This distance between the cover slip and the objective lens is your working distance. Scanner and LPO are
used within a greater working distance. Do the same then with LPO.
8. Lastly, swing the HPO in place. Always check your working distance. When using the HPO, use the fine adjustment
knob instead of the coarse adjustment knob. HPO is used within a short working distance meaning that the
objective is closer to the cover slip and almost touching. Again, use the fine adjustment knob to prevent
breakage.
9. Get the total magnification of your specimen observed under each objective by multiplying magnification of the
ocular with that magnification of the objective lens used.
10. Observe how the size of your specimen changes with each objective. Observe also the orientation or the position
of your specimen under the microscope. Why is this so?
11. To determine the size of an object observed under the microscope, one has to use the ocular micrometer, a
linear measuring device embedded on a piece of glass. But before this device is used, it has to be calibrated first.
Calibrating the ocular micrometer would need another measuring device, the stage micrometer.
12. Replace your regular eyepiece with an ocular micrometer. Mount the stage micrometer on the microscope stage.
Put the scanner into place. Peer then onto the ocular and locate the scales on the ocular micrometer and the stage
micrometer. Adjust the stage micrometer and the ocular so that their fist lines coincide with each other (Refer to
your instructor’s illustration/videos links).
13. Having done the previous step, find another two lines from both stage and ocular micrometer that coincide with
each other. Count then the number of lines or divisions found in between the overlapping or the coinciding lines
in the ocular and in the stage micrometer.
14. Having the number of lines determined, solve for the Calibration factor by dividing the number of lines/divisions
in the stage micrometer divided by the number of lines/division in the ocular micrometer multiplied by 0.01mm.
CALIBRATION FACTOR = # of lines or divisions in the stage micrometer in between coinciding lines (0.01mm)
# of lines or divisions in the ocular micrometer in between coinciding lines
15. Repeat step using the LPO and HPO.
Note: Every objective of every microscope to be used has to be calibrated first.
16. After you have calibrated your ocular micrometer, get your specimen’s measurement by counting the number of
divisions/lines in the ocular that covers it from side to side multiplied by the calibration factor solved earlier for
the specific objective used. Observe and measure your specimen using the scanner, LPO and HPO.

C. Determining Total Magnification:

OBJECTIVE LENSES – Notice the set of objective lenses on the revolving nosepiece. These lenses allow you to
change the degree of magnification. Some of our microscopes have four objective lenses while others have only
three. The degree of magnification for each objective lens is indicated on its side.

4X – This objective magnifies the image by a factor of 4. It is referred to as the “scanning objective” since it is
used to scan the slide to locate the specimen before viewing it at higher magnification. Your microscope may
not have this objective lens, in which case
you can begin with the 10X objective.
10X – This objective magnifies the image by a factor of 10 and is referred to as the “low power” objective.
40X – This objective magnifies the image by a factor of 40 and is referred to as the “high power” objective.
100X – This objective magnifies the image by a factor of 100. It is referred to as the “oil immersion objective”
since it requires a drop of immersion oil on the slide to provide good resolution. You will not be using this
objective lens.

Fill in the table below. (12 points)

Eyepiece Objective
Objective Lens X = Total Magnification
magnification magnification
Scanner X =
LPO X =
 HPO X =
OIO X =
D. Observing Slides (12 points)
1. From any resources, look for one specimen observed under the three objective lenses and draw their
differences in appearance under the microscope in the circles below. Put description/caption below and
solve for their total magnification power. [OL-objective lens: scanner, LPO or HPO; TMP-total mag.power]

OL: _________ TMP: _________ OL: _________ TMP: _________ OL: _________ TMP: _________

Specimen Description:

2. Describe the orientation/position of the specimen under the microscope? Why is this so?
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E. Classify the following part of the monocular microscope into mechanical, illuminating and magnifying
components. Write Mc if it is mechanical, Li if it is illuminating and Mg if it is magnifying. Write the answers on the
space provided before each number. (15 points)

________1.) mirror ________6.) coarse adjustment knob ________11.) base

________2.) condenser ________7.) revolving nosepiece ________12.) stage
________3.) eyepiece ________8.) oil immersion objective ________13.) HPO
________4.) draw tube ________9.) fine adjustment knob ________14.) arm
________5.) stage clip ________10.) scanning objective ________15.) LPO

F. Identify what part of the microscope is being described. (15 points)

______________________________1.) Reflects light

______________________________2.) Concentrates light
______________________________3.) Focuses the HPO or OIO
______________________________4.) Regulates the amount of light
______________________________5.) Holds the ocular or the eyepiece
______________________________6.) Facilitates the shifting of objectives
______________________________7.) Focuses with very low magnification
______________________________8.) Tilts the microscope when necessary
______________________________9.) Connects the base and the body tube
______________________________10.) Objective with the highest magnification
______________________________11.) Controls the opening of the iris diaphragm
______________________________12.) Keeps off dust particles from the objective lenses
______________________________13.) Lowers or raises the body tube in a faster manner
______________________________14.) Supports the whole microscope in an upright position
______________________________15.) Allow the passage of light from the objective to the ocular

G. ANALYSIS:
1. What did you notice that was the same when observing slides? (2 points)
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2. What happened when the magnification is changed from low to high power when observing these types of
cells? Use details when describing your observations. (3 points)
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3. What are some parts of the cell that you can possibly observe in high power? (2 points)
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4. State when do we need to use scanner, LPO, HPO and OIO? (3 points)
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5. What is numerical aperture? What is its relationship to magnification of the objective? What is its relationship
to the resolving power of the objective? (3 points)
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6. When do we need to calibrate the micrometer eyepiece? Why? (2 points)
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7. Cite some other types of biological microscopes and briefly discuss their specification. (at least 12 points)
8. List all reference materials used.