Name: ___________________________________________________ Rating ______________________

Yr/Section: ______________________________________________ Date Performed: ______________________

Date Submitted: ______________________

Activity No. ______

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
 Learn how to properly handle the microscope.
 Determine the total magnification of the microscope.
 Demonstrate the proper procedures used in correctly using the compound light 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
 Compound/Light microscope  Yeast cell
 Prepared slides  Glass slide and cover slip
 Letter e  Ocular and stage micrometer

A. Microscope Handling
1. Carry the microscope with both hands --- one on the arm and the other under the base of the
microscope.
2. Examine the microscope. Label the parts and give their functions.
B. PROCEDURE
1. Observe proper handling of the microscope. One hand on the arm while the other hand at the base.
2. Be acquainted with your microscope’s parts and functions.
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. Mount your letter “e” onto your slide and cover it with cover slip.
6. Mount your slide with letter “e” on the microscope stage. Secure your slide using the stage clips.
7. Observe your slide using 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 letter “e” changes with each objective. Observe also the orientation or the position
of your letter “e” under the microscope. Why is this so?
11. Be prepared to observe a live specimen. Observe a yeast cell.
12. 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.
13. 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).
14. Having done the procedure in #12, 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.
15. 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
16. Repeat step #12-14 using the LPO and HPO.
Note: Every objective of every microscope to be used has to be calibrated first.
17. After you have calibrated your ocular micrometer, observe a yeast cell. Describe its appearance, measure its cell
size. Get the cell’s measurement by counting the number of divisions/lines in the ocular that covers the cell from
side to side multiplied by the calibration factor solved earlier for the specific objective used. Observe and
measure the yeast cell 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.

Locate the numbers on the eyepiece and the indicated objective lens and fill in the table below.
Eyepiece Objective
Objective Lens X = Total Magnification
magnification magnification
Scanner X =
LPO X =
HPO X =
OIO X =
D. Observing Slides
1. Sketch the appearance of letter “e” under each objective. Put description/caption below.

OL: _________ TMP: _________ OL: _________ TMP: _________ OL: _________ TMP: _________
2. Describe the orientation/position of your letter “e” under the microscope? Why is this so?
___________________________________________________________________________________________
___________________________________________________________________________________________
3. What are the calibration factors of your objectives? Show your computations.
CFScanner=__________ CFLPO=__________ CFHPO=__________

4. Draw the yeast cell observed under each of the objectives.

OL: _________ TMP: _________ OL: _________ TMP: _________ OL: _________ TMP: _________
Size: ________ Size: ________ Size: ________
Computation: Computation: Computation:

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.

________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.

______________________________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. What happened when you changed the magnification from low to high power when observing these types of
cells? Use details when describing your observations.
___________________________________________________________________________________________________________________________________
___________________________________________________________________________________________________________________________________
___________________________________________________________________________________________________________________________________
3. What are some parts of the cell that you can observe on high power?
____________________________________________________________________________________________________________
4. State when do we need to use scanner, LPO, HPO and OIO?
___________________________________________________________________________________________________________________________________
___________________________________________________________________________________________________________________________________
___________________________________________________________________________________________________________________________________
___________________________________________________________________________________________________________________________________
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?
___________________________________________________________________________________________________________________________________
___________________________________________________________________________________________________________________________________
___________________________________________________________________________________________________________________________________
6. When do we need to calibrate the micrometer eyepiece? Why?
___________________________________________________________________________________________________________________________________
___________________________________________________________________________________________________________________________________
___________________________________________________________________________________________________________________________________

7. Cite some other types of biological microscopes and briefly discuss their specification.
8. List all reference materials used.
Name: ___________________________________________________ Rating ________________________________
Yr/Section: ______________________________________________ Date Performed: ______________________
Date Submitted: ______________________

Activity No. ______

Cell Structure

Label each structure for the plant and animal cell diagrams below:

Match each cell structure/organelle on the left with its function on the right:

____ nucleus A. modification, sorting of proteins

____ endoplasmic reticulum B. where cellular respiration occurs
____ Golgi apparatus C. gelatinous outer layer of prokaryotic cells
____ mitochondrion D. small structure that carries out protein synthesis
____ chloroplast E. projection used for motility
____ plasma membrane F. synthesis of lipids, secretory pathway proteins
____ cell wall G. stores water and other materials in plant cells
____ capsule H. contains the genetic material (DNA)
____ flagellum I. where photosynthesis occurs
____ ribosome J. provides external support/structure in some cells
____ centriole K. barrier between inside/outside of cell
____ central vacuole L. plays an important role in cell division

Place a check mark or “X” indicating a structure/organelle is present in the indicated cell type:

CELL nucleus ER Golgi mitochondrion chloroplast central plasma cell capsule

TYPE apparatus vacuole membrane wall
bacteria
plant
animal