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This activity gives the students the opportunity to use a

basic instrument of biology– the microscope. The microscope
enables the student to observe enlarged images of tiny objects.
There are various types of microscopes that have different
functions and levels of magnification. This activity will familiarize
MODULE 1 the students with the proper care and use of a brightfield
microscope, a typical compound light microscope.

Objectives

At the end of the activity, the student should be able to

PART A
1. identify the parts of a typical microscope and learn their
functions.
2. focus specimens under the compound light microscope.

THE Materials
MICROSCOPE Compound light microscope glass slides
newspaper cut-out coverslips

Procedure

A. The Microscope and its Parts

Locate the parts of the microscope listed below and label the
illustration in the Activity Sheet:

a. The eyepiece or ocular is the lens through which the

object or the specimen is viewed. This lens usually magnifies the
image passing through it by 10 times. It also has a pointer which
may be used to pinpoint the object or part of an object being
viewed.

b. The revolving nosepiece holds four objective lenses.

Grasp the nosepiece and note that it may be rotated, bringing
each objective lens into place. Feel the distinctive "click" as each
objective is in proper position for viewing.
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c. The objective lens gives the initial magnification. The microscope may have four
of these, located on the revolving nosepiece, enumerated below. In general, the
lower the magnification, the shorter is the objective lens and the farther it is
positioned from the specimen when focused.

- The scanning objective is the shortest objective which magnifies the size of an
object 4 times. It has the broadest field of view of all the lenses and is used for
initial focusing of objects.
- The low-power objective (LPO) magnifies by 10 times.
- The high-power objective (HPO) magnifies by 40 times. It is much longer than
the first two.
- The longest objective is the one with 100x magnification. This is the oil
immersion objective (OIO).

d. The stage is the flat surface with a round opening or aperture in the center. The
slide or the specimen to be viewed is positioned on the stage over the aperture.
The stage may be equipped with a mechanical device into which the slide is
clipped. This so-called mechanical stage allows the movement of the slide to be
manipulated by means of two knobs located underneath the stage. Other
microscopes do not have a mechanical stage and instead, stage clips are used to
fasten the slides on the stage.

e. The substage condenser is a lens located under the stage aperture, and its
distance from the stage is controlled by a knob. Its function is to concentrate the
light from the source and focus it on the specimen.

f. The iris diaphragm is beneath the condenser and functions to control the amount
of light passing through the specimen. This can be manipulated to affect a change
in light intensity.

g. The substage illuminator is the source of light which illuminates the object being
viewed. This may be replaced by a mirror.

h. The illuminator control knob is usually located on the side of the base i.e., the
part that supports the entire microscope. Also found just below the mechanical
stage adjustment knobs, it controls the light output of the illuminator.

i. The adjustment knobs are located on both sides of the microscope, just above
the base. There are two of these, usually with one smaller and located just outside
the other.

- The coarse adjustment knob is the bigger of the two. Its function is to focus
when using the scanning and low-power objectives.

- The fine adjustment knob is used in focusing at the high-power and oil-
immersion objectives.
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B. Magnification. The compound light microscope combines the magnifying power

of the eyepiece with that of the objective lens. The magnifying power is etched on
the housing of each lens. Simply multiply the magnification values of the objective
in place and the eyepiece to see how many times the specimen is enlarged.
Calculate the total magnification of the microscope with each objective. Write the
answer in the Activity Sheet.

C. Resolving Power. This is a measure of lens quality. Quality lenses have high
resolving power, the ability to deliver a clear image in fine detail. If the magnification
of the lens is high, but the resolution is low, it is of little value. Although such an
image would be enlarged, it would not be clear enough to show details. Resolution
is also affected by the cleanliness of the lens. It is a good practice to clean the
lenses with lens paper before and after every use.

D. Field of View. This is the size of the area that the lens views. The higher the
magnifying power of an objective lens, the smaller the area viewed. When one
switches to a lens with higher magnification, the central portion of what was visible
under low power is seen further enlarged. It is important to center the specimen
before increasing magnification, to prevent one from losing sight of the specimen.

E. Using the Microscope

1. Cut a small letter “e” from a newspaper article. Orient it upright and mount it on a
clean glass slide. Add a drop of water before placing a coverslip to secure it in
place. Be sure that no bubbles are formed.

2. Open the spring arm of the mechanical stage, place the slide in the proper position
on the stage and carefully return the spring arm to its closed position. This will hold
the slide in place. Using the control knobs, maneuver the slide on the stage so that
the tiny “e” is positioned over the stage aperture.

3. Illuminate the field of view by maneuvering the mirror or adjusting the light output
of the illuminator. Turn the revolving nosepiece so that the scanning objective
snaps into place directly over the aperture. Using the coarse adjustment knob
raise the stage so that the slide is as close to the objective as possible. While
viewing through the eyepiece, use the same knob to slowly lower the stage and
move the slide away from the objective until the letter “e” is in focus. Sketch a
picture of the letter “e” with the naked eye and its image as seen through the
microscope. Is there a difference? Using the mechanical stage, move the “e”
around. If the slide is moved to the right, what is the apparent direction of movement
of letter “e”? Now, carefully center the “e” in the field and proceed to the next step.
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4. Without making any other adjustment to the microscope, switch to the low power
objective. Focus the image using coarse adjustment screw and compare it to
the previous one. Is it still in the center of the field? If not, center it once more.

5. Again, without making other adjustments, rotate the revolving nosepiece to the
high-power objective. Viewing the objective from the side, note how close it is
to the slide. Look through the eyepiece, and using the fine adjustment knob,
bring the “e” into focus. If all is dark, manipulate the substage condenser and
iris diaphragm to improve the illumination of the specimen.

A feature of a good microscope is its parfocal capability. This means that when a
specimen is in focus under low-power magnification, one can switch to high-power
magnification and have the specimen remain in fairly good focus. Usually, a slight
adjustment with the fine adjustment knob is all that is required for a sharp image.

6. After focusing the letter “e”, put the scanning objective back into place and
remove the slide.

7. Obtain a slide of a protozoan (e.g., Paramecium, Amoeba) and practice focusing

the specimen using the compound light microscope. Draw the protozoan.

References:

Todd, JC. Clinical Diagnosis by Laboratory Methods. Philadelphia, PA, W.B. Saunders
Co.

Miller, Stephen A: General Zoology Laboratory Manual. 5th ed. McGraw-Hill.

http://webnt.calhoun.edu/distance/Internet/Natural/Biol103/New/HTML/Bio103/h.tips .

http://www.vscc.cc.tn.us/academic/math/biol/biol100/lab1.htm
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All living organisms are made up of cells that form

the basic unit of structure and function. The study of cells
constitutes the science of cytology. There are two basic
types of cells: prokaryotic cell, characterize by the
absence of a nucleus in which the genetic material is not
membrane-bound and the eukaryotic cell, characterize
by the presence of nucleus and membranous
organelles.

Cells differ in size and shape, which relate to their

structure and function, location in the body, and
relationship with the other cells. Under the compound
light microscope, three parts can be seen in the animal
PART B cell: the cell membrane, the structure that surrounds the
cell and regulates the flow of substances between the
cell and its surroundings; the nucleus, usually spherical
or ovoid structure that contains the genetic material; and
the cytoplasm, the substance outside the nucleus
where the other structures such as organelles are
ANIMAL CELL located. On the other hand, under the electron
STRUCTURE microscope, other parts of the cell can be seen, each
performing a specific function.

Objectives

At the end of the activity, the student should be able to:

1. identify the basic parts of a cell.

2. identify different cell shapes and relate it to the cell’s
function.

Materials

microscope coverslip
toothpick saline solution
methylene blue glass slide
cross section (x.s.) of stomach
x.s. frog's ovary medicine dropper
x.s. frog's testis hay
infusion frog’s blood smear
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Procedure

A. Preparing a wet mount of human cheek cells.

1. With the blunt end of a clean toothpick, gently scrape the inner cheek lining.

2. Thinly disperse the scrapings in a drop of normal saline solution (0.9% NaCl
solution) on a clean glass slide.

3. Get a cover slip, put one edge of the cover slip on one side and slowly lower it over
the drop of saline solution.

4. Examine the slide first under the LPO, then under the HPO. Locate the cheek cells.
Identify the shape and the distinct parts of the cell.

5. Remove the slide from the stage.

6. Repeat the same procedure but use 1-2 drops of diluted methylene blue stain
instead of normal saline solution.

7. Focus the stained preparation under both LPO and HPO. Take note of the
difference between unstained and stained preparation.

8. Draw and label the image of the cheek cells in the Activity Sheet.

B. Shapes of Animal Cells

Cells show complementarities of shape and function. The shape is also affected
by the location of a cell and its relationship with other cells.

1. Examine a prepared slide of a cross-section of the stomach. Focus on the muscular

layer of the organ and look for cells that are fusiform or spindle-shaped. These are
smooth muscle cells. Label the photomicrograph in the activity sheet.

2. Examine a prepared slide of the frog’s ovary. Note the spherical-shaped eggs
scattered in the section. Focus an isolated spherical cell with a distinct nucleus.
What structure is very evident inside the nucleus? Label the photomicrograph in the
activity sheet.

3. Examine a prepared slide of the frog’s blood smear. Focus cells that are oval in
shape and with a distinct nucleus at the center. These are the red blood cells.
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4. Using the same slide as above, look for cells that are smaller than red blood cells.
They are less numerous and are nucleated. They have no definite shape and hence
are said to be amorphous. These are white blood cells. Identify both types of blood
cells and label the photomicrograph in the Activity Sheet.

5. Examine the frog’s testis under HPO. Focus the sperm cells/spermatozoa. Note the
whip-like flagellum that gives the cell a threadlike appearance. Label the
photomicrograph in the activity sheet.

C. Examination of Live Cells

1. Obtain a hay infusion. It is a culture of microorganisms made by adding hay or cut

grass in a jar with water collected from a pond or similar body of water. After a while,
the water turns cloudy due to the increase in population of the microorganisms
present in the jar.

2. Using a medicine dropper, obtain water samples at certain levels (top, middle, or
bottom) of the jar and put a few drops on a clean glass slide. Place a coverslip and
view using the low-power objective of the microscope. Draw examples of the live
microorganisms seen in the slide.

References

Warren, D. Dolphin: Biology Laboratory Manual. 4th edition. Copyright 1997. The
McGraw-Hill Co., Inc.

Crescencia, N. Catada et al. Laboratory Manual for General Zoology. Copyright 1979.
Vibal Publishing House, Inc.