BIO 1041 INTRODUCTORY BIOLOGY LABORATORY

Laboratory # 1. Microscope Lab

Psalm 104: 25. There is the sea, vast and spacious, teeming with creatures beyond number-living
things both large and small.

Read through the lab and answer questions before you leave lab. Answers can be discussed dur-
ing lab class, so that you can use them to study for the exam next week. Use the back of this
handout for drawings, if necessary. Number and label all figures.

I. Why do we use microscopes?

A. Find a personal object in your pocket or purse. Pick up a glass container (flask or beaker),
place a specimen on the other side (behind the flask). Look through the container at your speci-
men. Add water from the sink to the container. Now look at your specimen through the container
with water in it.
· Q1. Did magnification of your specimen take place? YES
· Q2. Through which container, with or without water? FLASK OF 100MIL , WITH WA-
TER.
B. Pick up a hand lens (looking glass or magnifying glass). Examine your specimen.
· Q3. Which of the three microscopes used magnified the most (container, container with
water, hand lens)? HAND LENS
1. Draw the specimen, as you see it through the lens, in a rough sketch.

II. Scientists before the 1600's noticed that objects sitting behind containers of water were mag-
nified when the sun shined into the container. In the 1600's a Dutch lens grinder and cloth mer-
chant, Anton van Leewenhoek, built one of the first microscopes. This primitive device used a
magnifying glass to make the specimens living in ponds, soil and even on his own body, seem
larger.

By the 1700's, Robert Hooke, an English scientist, was examining a cork (oak) that was used to
seal a bottle. He saw little cavities separated by walls in the cork, as well as in other plant tissues.
He called each cavity surrounded by a wall a cell. Hooke constructed his own magnifying de-
vice, based on that developed by van Leewenhoek.

There are a variety of microscope models used for industry, the medical profession and educa-
tional purposes. Many microscopes are adaptations of the simple compound light microscope

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(CLM). Manipulation of different light sources confers each type of CLM microscope specific
functions. Other well known microscopes are the stereo or dissecting microscope and the elec-
tron microscope. The dissecting microscope is used to examine large specimens or to perform
dissections. Electron microscopes magnify tiny specially prepared specimens thousands of times
their size.

III. The Compound Light Microscope (CLM): This microscope is usually used to examine
only tiny or thin specimens on slides, such as microscopic organisms or tissue sections.
See Figure 1.

Figure 1: Compound light microscope (CLM)

Label the parts: A- eye piece/ocular
B- Body tube
C- Revolving nose piece.
D- Objectives (this has 4)
E- Arm
F- Stage
G- The condenser
H- Iris diaphragm
I- Coarse Adjustment
J- Fine Adjustment
K- Light Source
L- Base

A. Find the part of the microscope that you look through. It is called an eye-piece or ocular.
It has a magnifying power of 10X (which you will normally see written on it in white letters).
If your microscope has one eye-piece it is called monocular, and if it has two eye-pieces it is
called binocular.
· Q4. Is your microscope monocular or binocular? BINOCULAR

B. Look for the black, square stage in the middle of the microscope. The stage is a base for the
slide containing your specimen. It holds the slide securely inside metal clips. Use a prepared
slide with the letter "e" on it. Place it on the stage, using the clip holders. Look under the stage
for two gears (slide movers). Play with the gears very gently, and use them to move the slide
side-to side,
and back and forth.

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· Q5. When you move the slide to the left, in which direction does the specimen appear to
move? (left or right) : RIGHT
· Q6. When you move the slide away from you, in which direction does the specimen appear
to move (towards or away from you)? AWAY
C. There is a revolving nose-piece above the stage, held by the microscope body, that contains
three or four tubes. These are called objectives and are used to make the specimen appear larger
than it is. Each one has writing on it indicating a specific power of magnification. Most of the
microscopes at PBA have the same four powers. On some microscopes an objective will be la-
beled oil immersion. This objective is only used to highly magnify specimens such as bacteria.
After a drop of oil is added to the slide, the specimen is focused by gently lowering the objective
into the oil. After viewing, oil should be cleaned from the objective and from the slide with tis-
sue paper.
· Q7. What are the 4 magnifying powers on your microscope objectives? (Look for an "X"
after the number) 4,10,20,100

D. To focus a specimen, rotate an objective by moving it gently and clicking it into place. Click
the lowest objective, 4X, into place using your slide moving gears. There is a circle around the
letter "e" on the slide. That circle should be on top of the hole in the center of the stage. Now
look for two knobs (one on top of the other) on the arm, but below the stage. These are called fo-
cusing knobs. The coarse adjustment moves the stage up and down to focus, while the fine fo-
cus precisely focuses on the specimen. Everything that you see through the objective is called the
field of view.
· Q8. Which knob controls the fine adjustment? (the large knob or the small knob) SMALL
NOB

E. A very important part of focusing your specimen involves the use of light. The inside of your
microscope consists of a series of prisms that reflect light with which to view your specimen.
You must be careful to hold the microscope upright when moving it, and never slide it around on
the desk sideways. Directly underneath the stage is a light source. The light is focused by a con-
denser, through an opening, upwards through your specimen. The opening is controlled by the
iris diaphragm lever. It moves from side-to side. Return to the slide of the letter "e". Examine it
using the 4X objective. Move the lever to allow the most light to pass through the specimen.
Now, move the lever in the opposite direction, to allow the least light to pass through the speci-
men.
· Q9. Is the letter "e" clearer to see when the most light possible is focused on it, or when
the least light is focused on it? MOST LIGHT
· Q10. What would happen if strong light is focused on a specimen that was transparent (as
are most living cells?) INVISIBLE. LOOSES DEFINITION.

F. To calculate the total magnification of your specimen, multiply the power of the eye-piece
against the power of the objective that you are using. The power of the eye-piece is always 10X

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on microscopes at PBA. Using the slide of the letter "e", now on the stage, change the objectives,
focus the specimen and fill in the blanks in the following table.

· Q11. ocular objective total magnification

10X 4X 40X (example)
10X 10X 100X
10X 75X 750X
10X 100X 1000X

G. The main purpose of using a microscope is to see the specimen as clearly as possible under
conditions that appear to enlarge it. When the specimen is too small, human eyes lose the ability
to see the specimen clearly (resolve details). Microscopists call this depth of field. Depth of field
becomes smaller at higher magnification, since fewer layers of the specimen can be seen. Focus
the letter "e" at each of four objectives.
· Q12. At which objective can the largest portion of the letter be seen? 4
· Q13. At which objective can the least portion of the letter be seen? 100 OIL
· Q14. Does depth of field increase or decrease when the smallest portion of the letter can be
seen? INCREASE

To further illustrate depth of field, use a prepared slide of three threads. Focus the threads at
each of four objectives.
· Q15. Name the highest objective at which each color be distinguished from the others? 10
· Q16. At which objective do colors blend together?_40 AND 100

IV. Stereo or Dissecting Microscope . This type of microscope has much lower magnification
than the CLM. It may be used to study larger specimens, and dissections may be performed on
its stage. See Figure 2.

Figure 2: Dissecting Microscope

Label the parts A- Eye piece/ocular
B- ARM
C- FINE ADJUSTM
D- BODY TUBE
E- OBJECTIVE
F- LIGHT
G- STAGE

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 H- BASE

A. Study the dissecting microscope. This microscope can also be monocular or binocular.
Identify the oculars. They magnify the specimen 10X.

B. Identify the stage area. Compare the distance of objectives to the stage on the dissecting mi-
croscope with that of the CLM.

C. There are usually one or two objectives, housed inside a large tube. Look on the side of the
microscope or the large housing and find the magnification of the objectives.
· Q17. If there is no identification of magnification written on the microscope parts, how
could you tell whether you were using a low or a high magnification?
COLOR OF TUBE. THE CLOSENESS OF THE STAGE. DEPTH OF FIELD INCREASE.

D. There are either one or two focusing knobs, and one light source. Pick out a specimen from
your belongings or the prep table and examine it using all objectives on your microscope.
· Q18. What are the powers of the objectives? 4,10,40,100
· Q19. What benefits do microscopes offer in viewing tiny specimens, rather than using wa-
ter or hand lenses?_CONTROL OVER VARIETY OF DEPTH, LIGHT, MAINTAINS
ORIGINAL ENVIRONMENT FOR THE SPECIMEN
· Q20. Which microscopes have you heard of that magnify specimens hundreds of thou-
sands of times more than compound light microscopes?ELECTION MICROSCOPE