Laboratory 1-2

Microscope and the Cell

Full Name

Required materials:
Lab coat paper lab coats are fine
This lab manual and a pen
Long pants and closed shoes

Provided materials:

Light microscope
Microscope slides and cover slips
Newspaper
Sterile toothpicks
Saline solution
Methylene blue dye
Prepared slides with seven different tissues: (1) regular connective, (2) striated muscle,
(3) columnar epithelial, (4) bone, (5) squamous epithelial, (6) adipose, and (7)
neurons

Objectives:

Examine several different objects through the microscope and become familiar with the
proper use of the instrument.
Be able to locate and give the function of each of the following parts of the microscope.
Identify the basic parts of animal cells while viewing them with a microscope.

Light Microscope Rules

Microscopes are expensive to replace. Currently they are going for about $1,500 used on
eBay. So, in order to maintain their quality, you must be particularly careful when handling
them and follow these rules:
1. Always carry microscope with TWO hands. Hold it upright so that the ocular lenses do
not slip out.
2. When finished viewing, use the coarse focusing knob to move the stage down, away from
the lens.
3. Always use the provided lens paper to clean the lenses. This avoids scratching the glass.

Never touch any lens with your fingers or anything else except lens paper.

NEVER use Kimwipes, paper towels or Kleenex.

1. Always begin with LOWEST POWER lens to focus on your sample, then work up in
magnification from there.

Never use the coarse knob at high power, or you may crash the lens into your
slide.

1. TURN OFF the light when not in use to avoid burning out the bulb and drying out your

sample.
2. RETURN the scope to the cabinet CLEAN (no oil or water or slide) and with the LOW
POWER objective facing the stage. Each student is assigned one particular
microscope for use throughout the semester; the microscopes will be inspected
regularly for cleanliness, and points will be deducted if your microscope is found to
have oil on the lenses.
3. If the ocular lenses (eyepieces) must be removed to fit the scope into the cabinet, please
replace the black plastic caps on the eye tubes to keep them dust-free (they are very
difficult to clean out). Take care not to misplace the caps while using or transporting
the scope.
4. Always clean the immersion oil off of the 100x objective after use (and any other part of
the microscope that it spilled onto) using lens paper and a little bit of xylol solution.
If the oil dries on the lenses, they will become cloudy and very difficult, if not
impossible, to clean. Check the 40x lens for oil contamination also.

Components of Light Microscope

The microscope is the basic instrument for studying things too small to be seen with the
unaided eye. The microscope has opened more horizons to mankind than the telescope. It
has allowed us to explore the worlds of miniature life. The microscope has helped humans
to explore the inner world of tissues and cells. By using various dyes that attach to different
kinds of chemicals, the microscope even allows us to look at the biochemistry of different
parts of the cell. For example, we knew from the microscope that the nucleus was rich in
DNA and proteins long before Watson and Crick broke the genetic code. The light
microscope alerted us to the presence of ribosomes (RNA-containing structures) in the
cytoplasm long before technology allowed us to photograph them with the electron
microscope. Good microscopists drew organelles within the cytoplasm long before
scientists understood the functions of these structures.
There are many different kinds of optical (or light) microscopes, but they all have certain
parts in common. Locate the following parts of the microscope:
Arm: This is the curved portion of the instrument that supports the optical portion of the
microscope.
Base: This is the heavy bottom portion of the instrument that gives it stability.
Ocular lens or eyepiece: (the lens closest to your eye) This is also known as the eye
piece. It consists of several pieces of glass that have the effect of magnifying objects
by a factor of 10X. Many ocular lenses have a pointer mounted in them.
Objective lenses: The lens closest to the object you are viewing, this is the main (and
most expensive) lens.

Revolving nose plate: This portion of the microscope allows different objective
lenses to be rotated into position.

Low-Power objective lens: These lenses are located on the nose plate and have
the ability to magnify objects by a factor of 4 or a factor of 10.

High-Power objective lens: (high dry): This lens is located on the nose plate
and has the ability to magnify objects 40 times. (Your microscope may have
a high- power objective that magnifies by a slightly different amount.)

Oil immersion objective lens: This lens is located on the nose plate and has the

ability to magnify objects 100 times. Immersion oil is always used between
the object (microscope slide) and this lens. (WHY?)
Sample loading area:

Stage: This is the flat platform upon which slides are placed. A mechanical stage
has a device attached to hold the slide and move it back and forth or up and
down in the field of view using knobs.

Stage clips: These structures are used to hold the slide in place on the stage
(Caution: Some microscopes are fitted with mechanical stages that do not
have clips. These models have spring-loaded devices that hold the slide in
place while allowing it to be moved on the stage. Your instructor will show
you how to use this mechanism.)
Focusing knobs:

Coarse focus knob: This is the large knob at the base of the microscope that
varies the distance between the objective lens and the stage. A small change
in the position of the knob causes a large change in the distance between the
stage and the lens. Use this knob to focus when using low power objectives.

Fine focus knob: This is the smaller, inner knob. A large change in the position
of the knob changes the distance between the objective lens and the stage
very little. Use this knob to focus when using high power objectives.
Diaphragm or light aperture: This part is located below the stage and regulates the
amount of light that passes through the opening in the stage. A small lever is used to
change the size of the opening through the light aperture. When using low power
objectives, the iris should be partially closed to enhance the definition and detail of
the image.
Lamp: This is located below the stage and supplies the light necessary for viewing
objects.
On/off Switch: This is located on the lamp (or near it) and is used for turning the lamp
on and off.

Figure 1: Components of a light microscope.

I. Match the components below with the corresponding letter above.

1. Arm

2. Fine focus knob

3. Coarse focus knob

4. Base

5. High-power lens

6. Light aperture/diaphragm

7. Lamp

8. Low-power lens

9. Ocular lens/eye piece

10. Stage clips

12. Revolving nose plate

11. Stage/on-off switch

II. Magnification
The compound microscope combines the magnifying power of the ocular lens with the
magnifying power of the objective lens. The magnifying power of each lens is marked on
the outside of the lens itself. Simply multiply the magnification value that is marked on the
ocular lens by the value marked on the objective lens to determine how many times the
image of the object has been increased.
Notice that your ocular lens magnification is 10X. Since the low-power objective lens is also
marked 4X, the total low-power magnification is 10X times 4X = 400x.
Microscopes often have other objective lenses (high-dry, oil immersion) with other
magnifying powers. Calculate the total magnifying power of each ocular lens/objective lens
system.
Lens name
Low power lens

Ocular lens
magnification
10X

Objective lens
magnification
4X

Total
magnification
40X

Field of View
You have already learned that different lenses have different magnifying powers. It is also
important to understand that each lens has a specific area that can be observed through it.
The higher the magnifying power of an objective lens, the smaller the area viewed. This is
sometimes hard to appreciate since the size of the circle of light you observe through the
ocular lens looks the same. However, when you switch from low power to high power, you
are only looking at the central portion of the low power field of view. Therefore, it is
important to center an object in the low-power field of view before you switch to high
power.

III. Viewing a newspaper letter

1. Make a wet-mount slide of a one-centimeter-by-one-centimeter piece of newsprint. Use
the smallest print available. Place the newsprint on the slide and place two drops of
water on top of it. Place a coverslip over the water and the newsprint.
2. Center the newsprint over the hole in the stage of your microscope.
3. Rotate the lowest power (4X objective) lens into position over the specimen. Make the
distance between the lens and the specimen as small as possible by using the coarse
focus knob.
4. While looking through the ocular lens, use the coarse focus knob to move the stage away
from the objective lens. You should eventually see some of the fibers in the paper
and perhaps a portion of a letter or two. If too much light is entering the lens, close
the light aperture and/or adjust the light intensity using the light control on the base.
5. Sharpen the focus by using the fine focus knob.
6. Adjust the light by manipulating the light aperture and light intensity control.

7. Center a letter in the low-power (4x objective lens) field of view. Focus to make the image
sharp. What do you see now?

8. Sketch the entire view of that you can see in the space below. If only part of a letter is
visible at one time, sketch only that part of the letter.

9. When you move the slide to the right, in what direction does the object appear to move
under the microscope? Circle what you found to be the case:
Left
10.

Up

Down

When you move the slide up, in what direction does the object appear to move under
the microscope? Circle what you found to be the case:
Left

11.

Right

Right

Up

Down

Without adjusting the focus, switch to a higher power (10X objective). You will now
be able to see only a smaller portion of the previous low-power field of view. Draw
a sketch of the new visible area in the space below. Be careful, you can drive the
lens through the slide. At this point, only use the fine focus knob to adjust the focus.

12.

You may now increase the power again- switch to the 40X objective. Use the fine
focus again and draw the final sketch in the space below.

13.

Note: If you have difficulty with any of these procedures, ask your instructor for
help.

Remember, when you are finished viewing a particular slide, always:

A)

Swing the high power lens out of the way and put the lowest power lens in
position.

B)

Use the coarse focusing knob to move the stage down, away from the lens.

C)

Use lens paper to clean immersion oil from the 100X oil immersion lens (and any
other lenses it may have contacted).

D)

Use Kimwipes to clean any water or immersion oil from the stage and from the
microscope slides if they are to be saved.

E)

Turn off the light.

F)

If you are finished for the day, unplug the microscope and wrap the cord around
the base, as you found it.

Structure of Cells
Two distinct types of cells exist in the biotic world. Organisms such as us, are composed of
cells that have (1) a nucleus, (2) chromosomes composed of DNA and protein, and (3)
membranous cytoplasmic organelles. Cells from these organisms are called eukaryotic cells,
meaning that they have a true nucleus. Prokaryotic cells lack the three characteristics listed
above. Prokaryotes are the bacteria and archea of the world (they are given this name
because they evolved before the nucleus was devised (pro- = before and -karyotic =
nucleus)). In this exercise, you will study the structure of only eukaryotic cells.
A cell is composed of two major parts: (1) a nucleus and (2) the cytoplasm. The nucleus is
always embedded in the cytoplasm. As you study the descriptions of these basic
components of cells, refer to the list below and Figure 2. Note that the figure contains more
detail than can be seen with an optical, or light, microscope. In fact, the organelles in Figure
2 were determined with an electron microscope capable going to 100,000X magnification.
The structure and function of the major organelles are described below:

Figure 2: The generalized animal (eukaryotic) cell

IV. Human Epithelial Cells

The epithelial cells lining the inside of your mouth are easily obtained for study. They
exhibit the basic characteristics of animal cells.
1. Prepare a wet-mount slide of human epithelial cells.
A)

Wash your hands thoroughly.

B)

With a sterile toothpick, gently (!) scrape the inside of your cheek.

C)

Gently swipe this toothpick across the middle of the slide.

D)

Add a drop of 0.01% methylene blue to the microscope slide.

E)

Cover the drop of fluid with a coverslip.

2. Observe the cells at both 100X and 400X magnification. Draw a diagram of your cells
at 100x and 400x magnification. Draw in the space provided below. Make sure your
diagram is large enough to include all that you see. Label the structures that you
can identify. Make sure you indicate the total magnification.
Note: You may need to reduce the light by closing the light aperture. Too much light is
often a problem in microscopy.

3. Estimate the size of both whole cell and its organelles using the ruler in the eyepiece.
Example: Each small tick in the eyepiece ruler is 1 millimeter (1 mm) and the large
numbered ticks are 1 centimeter (1 cm). In the figure below, we see an object that is 10.5
millimeters long in the microscope. The magnification is 100X. To get the actual length of
the object, we must divide by the magnification. 10.5 mm / 100X = 0.105 mm or 105 m
(micrometers).

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