ACTIVITY 1

MICROSCOPE AS A BIOLOGICAL TOOL

The microscope is one of the principal tools used in Biology. It was invented
through the efforts of the Dutch scientist, Anton Van Leeuwenhoek. In the laboratory,
the microscope serves as a very useful tool to help you discover the fascinating secret
of the living world, which the unaided eye cannot see. In this exercise, you will study the
proper use and care of this delicate instrument.

Objectives:
1. Identify the parts the compound microscope.
2. Describe the function of each part.
3. Manipulate the different parts of microscope correctly
4. Compute the magnification of the image and the drawing , and
5. Differentiate the objectives in relation to the size of the object.

Materials:
Compound microscope medicine dropper
Glass slide transparent or plastic ruler
Cover slip small letter “e” cut from an old news paper
Needle dissecting needle

Procedure:

Before studying the different parts of the microscope, bear in mind the following
points.

1. Other students from other classes also use microscopes. Always inspect it before
using and report to the instructor for any damage.
2. The microscope is delicate and expensive instrument proper care should be
exercised so other students can use it.
3. Manipulate the microscope only when you are thoroughly familiar with it.
A. IDENTIFICATION OF THE PARTS OF THE MICROSCOPE

1. Using figure 2, identify the numbered structures of the compound light

microscope described below and enter your answers in table 1.

Compound Light Microscope

Compound light microscopes are microscopes which use a combination of 2 or more

lenses to magnify the specimen and light (wether natural or electric) to view it. Below
are the parts of a compound microscope that were grouped into 3; the 1. Mechanical
parts, 2. Optical parts and 3. Illuminating parts. Familiarize yourself with these structure
as well as its function.
Mechanical Part
Diopter Adjustment – usually found in binocular type of microscope and is useful as a
means to change focus on one eyepiece so as to correct for any difference in vision
between your two eyes.
Draw Tube – a small tube attached above the body tube wherein the ocular or the
eyepiece is placed
Body Tube/ Head – a hollow tube through which light travels from the
objective to the ocular. separates the objective and the eyepiece and assures
continuous alignment of the optics. Also known as the Head, this structure carries the
optical parts in the upper part of the microscope.
Revolving nose piece/Turret – this part of the microscope can be rotatted and contains
3-4 objective lenses with varying magnifying power. The rotating ability of this part
facilitate the changing of lenses for viewing.
Dust Shield – the circular disc above the revolving nosepiece that protects the
objectives from dust particles
Arm/ Frame – supports and connects the body tube to the base of the microscope. It
also primarily serves as a means for carrying the microscope
Stage/ Mechanical Stage – the part where the object to be examined is placed. Stages
in some microscope are often equipped with mechanical ( thus the name mechanical
stage) device that holds the specimen slide in place and can smoothly move the slide
back and forth as well as from side to side.
Stage Clips/ Mechanical Clips – holds the slide securely in position.
Adjustment Knobs - the wheels that raise or lower the tube or stage of the microscope.
a. coarse adjustment – changes the elevation of the tube or stage rapidly for
preliminary focusing
b. fine adjustment – permits gradual changes in the elevation of the tube or stage for
final and accurate focusing
Inclination joint/ Hinge – A joint at which the arm is attached to the pillar of the
microscope.
This part allows tilting of the microscope at a desired angle.
Pillar –the part that extends upwards from the base to support all other parts, and to
hold the stage. It provides support to all the remaining parts of the microcope.
Base – what the microscope stand on and the part that supports the entire microscope

Optical Parts
Ocular/ Eyepiece/s – the part where you look into when viewing specimens under the
microscope. It usually contains 5X and 15X magnification or power lens and functions to
gather light and enhance magnification of specimens being viewed.
Objectives - refer to the cylindrical tubes attached to the revolving nosepiece for initial
magnification of the image. Each objective lens is color coded, this helps identify their
magnifying power.( Fig.2)
a.Scanning Power Objective- the shortest tube lens with the red band provides the
lowest magnification power (4x) of all objective lenses. It provides the observer
with enough magnification to go over the entire slide, essentially a “scan” of the
slide.
b.Low Power Objective – possesses the yellow band, magnifies 10x and allows
you to quickly scan a large area of the specimen, and to locate those areas which
need closer study with a high power objective.
c. High Power Objective – possesses the blue band and magnifies 40x, it is ideal
for observing very fine detail
 d. Oil Immersion Objective – are generally available from 40 to 120x. obtains
greater magnification than HPO. In using this a drop of oil is placed on the slide for
better refraction thus the name. It is also called the “wet” objective and has a white
band.

Fig.1 . Compound Microscope Objective lenses

Image Source: https://www.amazon.com/OMAX-Achromatic-Microscope-Objective-Coating/dp/
B00FG8ETY4

Illuminating Parts
Mirror/Illumination- it is used to reflect light from an external light source up through the
bottom of the stage. One side of the mirror is plain (used with natural light);the other is
concave (used with artificial light). Some microscope do not have mirror instead it has a
built in light source.
Irish Diaphragm – located above the condenser and below the stage and controls the
amount of light reaching the specimen.
Condenser – this lens system is located immediately under the stage and focuses the
light on the specimen.. Condensers are valuable with high power objectives and oil-
immersion lenses beacuse it focuses light to the specimen.
 Figure2. parts of the compound light microscope

B. MICROSCOPE MANIPULATION

1. After familiarizing each part of the microscope, proceed to the actual microscope
manipulation. But before you proceed, here are some reminders each time you
prepare for some microscopic work.

a. Make certain that the low power objective or the scanning objective is
present and in position. (A slight “click” will indicate that the LPO is already in
place.
b. Adjust the diaphragm to its large opening.
c. Look into the eyepiece and adjust the angle of the mirror to obtain good
illumination.
2. Cut a small letter “e” and place a drop of water on the center of the slide. In an
upright position, place the cut letter “e” on the droplet of water. Cover the droplet
with a cover slip carefully letting one side of the cover slip touch the droplet and
slowly lowering the other side using a dissecting needle. Be sure that no air
bubbles are trapped inside.
3. Place the slide on the stage of microscope with the letter “e” at the center of the
stage aperture. Secure the slides into place with the stage clips.
4. Now, slowly turn the coarse adjustment knob downward to lower the body tube
until the tip of the LPO almost touches the slide. While looking into the ocular,
slowly turn the adjustment knob upward to raise the LPO until the objects comes
into focus. Turn the fine adjustment knob to obtain a sharp image of the letter.
a. Draw the letter “e” in its actual size on the glass slide and the letter “e” as
seen under LPO of the microscope in worksheet 1.
5. While looking in the ocular, move the slide slightly to your right. Next move it to
your left.
a. In which direction does the image appear to move in each case?
Move the slides towards you. Move it away from you. (1).
b. In which direction does the image appear to move in each case?
6. Rotate the nosepiece to put HPO into position. Make sure the objectives moves
into position without hitting the stage or the slide. Listen for the click sound which
indicates that the objective is in place.
7. The microscope today are parfocal. This means that once image is in sharp
focus under LPO, it remains focus under HPO without further adjustments.
When at first try you fail to see the image under HPO, refocus under LPO then
repeat the procedure for focus with HPO until you see the image.

Note: do not use coarse adjustment knob when the HPO is in place. The reason
for this is may be obvious if you look at the distance between HPO and the top
of the slide. The distance is so close that a downward movement of the
objectives has a danger of hitting and breaking the slide. With the fine
adjustment knob, the downward and upward movement of the objectives are
very slight and do not normally hit the slide or stage.

Draw the letter “e” as seen under HPO in worksheet no. 1.

C. MICROSCOPE MANIPULATION

8. After familiarizing each part of the microscope, proceed to the actual microscope
manipulation. But before you proceed, here are some reminders each time you
prepare for some microscopic work.

d. Make certain that the low power objective or the scanning objective is
present and in position. (A slight “click” will indicate that the LPO is already in
place.
e. Adjust the diaphragm to its large opening.
f. Look into the eyepiece and adjust the angle of the mirror to obtain good
illumination.
9. Cut a small letter “e” and place a drop of water on the center of the slide. In an
upright position, place the cut letter “e” on the droplet of water. Cover the droplet
with a cover slip carefully letting one side of the cover slip touch the droplet and
slowly lowering the other side using a dissecting needle. Be sure that no air
bubbles are trapped inside.
10. Place the slide on the stage of microscope with the letter “e” at the center of the
stage aperture. Secure the slides into place with the stage clips.
11. Now, slowly turn the coarse adjustment knob downward to lower the body tube
until the tip of the LPO almost touches the slide. While looking into the ocular,
slowly turn the adjustment knob upward to raise the LPO until the objects comes
into focus. Turn the fine adjustment knob to obtain a sharp image of the letter.
b. Draw the letter “e” in its actual size on the glass slide and the letter “e” as
seen under LPO of the microscope in worksheet 1.
12. While looking in the ocular, move the slide slightly to your right. Next move it to
your left.
c. In which direction does the image appear to move in each case?
Move the slides towards you. Move it away from you. (1).
d. In which direction does the image appear to move in each case?
13. Rotate the nosepiece to put HPO into position. Make sure the objectives moves
into position without hitting the stage or the slide. Listen for the click sound which
indicates that the objective is in place.
14. The microscope today are parfocal. This means that once image is in sharp
focus under LPO, it remains focus under HPO without further adjustments.
When at first try you fail to see the image under HPO, refocus under LPO then
repeat the procedure for focus with HPO until you see the image.

Note: do not use coarse adjustment knob when the HPO is in place. The reason
for this is may be obvious if you look at the distance between HPO and the top
 of the slide. The distance is so close that a downward movement of the
objectives has a danger of hitting and breaking the slide. With the fine
adjustment knob, the downward and upward movement of the objectives are
very slight and do not normally hit the slide or stage.

Draw the letter “e” as seen under HPO in worksheet no. 1.

D. MAGNIFICATION OF THE IMAGE

The magnifying power of an objective is usually marked on its side as 5x, 10x
or 40x. If the objective is marked 40x, it means that the lens in the objective
forms an image that is 40 times larger than the specimen or object. If the ocular
is marked 10x, it means that the ocular magnifies the object 10 times. The total
magnification (M total) of the image you see in the microscope is obtained by
multiplying the magnifying power of the ocular (M objectives) with objective in
use.
FORMULA:
MTOTAL= (Mocular) (MObjectives)

Calculate the total magnification of image under different objectives (a).

scanner (b). LPO (c) HPO and enter the values in Table 2.

E. FIELD OF VISION

The field of vision is that circular lighted field where you can see the image of
the object or specimen.
1. Focus again the letter “e” at the LPO and HPO and then compare the
brightness of the field of vision.
2. Remove your wet mounted slide letter “e” and place a clear plastic ruler on the
stage and focus on the millimeter scale with the scanning objective. While looking
through the ocular, carefully, move the ruler until one of the vertical lines of the
millimeter scale is in line with the left boundary of the field of vision.

a. What you observe in the appearance of the line the center of the next line is
one millimeter. Count the number of the millimeter covered from left to right
side of the field that coincides exactly with the line in the ruler. Estimate the
fraction of the millimeter, which is included in the field. Estimate the
diameter of the 4x objective, 10x objective, and 40x objective field of vision
in millimeter (mm) and enter the values in table 3.
3. The area of the field of vision can be calculated by using the Formula A= 𝜋r2,
where 𝜋= 3.1416, and r=1/2 of the diameter of the field of vision or radius.
Compute for the area of the field of vision with 4x, 10x, and 40x objectives.
Enter the values in table 3.

a. What is the effect of changing from LPO to HPO on the area of the field of
vision? (3)____________
b. Where do you see most of the object, under LPO or HPO? Why?
(4)___________

F. CARE OF THE MICROSCOPE

1. The scanning objective should be in place.

2. The microscope must be clean.
3. The stage clips must be pointing forward.
4. The mirror should be in its vertical position.
5. The parts of the microscope with one hand grasping the arm of the
microscope and the other hand supporting the base.
 Activity No. 1
The Microscope as a Biological Tool

Group Number _____ Date Performed: _________

Members: __________________ Date Submitted: __________

__________________
__________________

A. Identification of the parts of the microscope

Fill in Table below with the parts and function of the compound microscope
below.
 Table 1 Parts of the Compound Light Microscope
Numbered Microscope Part Functions
Structure
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
B. Microscope Manipulation

Letter “e” as seen under LPO

1. A
Letter “e” in its actual size and
position on the slide n ___x s.
_ _
_____________________________________________________________
_____________________________________________________________
_____________________________________________________________
_______________________________________________________

2. Ans._________________________________________________________
_____________________________________________________________
_____________________________________________________________
_____________________________________________________________

Letter “e” as seen under HPO

___x
 C. Determination of the Total Magnification of the Image
Table 2. Magnification of the image under different objectives

Objective Magnifying Power Magnifying Power Total

the Objectives of the Ocular magnification of
the Image
Scanner
Low Power
Objective
High Power
Objective

D. Field of Vision

Table 3. Area of field vision

VALUES 4x 10x 40x

Diameter in (mm)
Area in square mm

Questions:
1. What other instruments are used by the scientist to observe object which cannot be seen
by the naked eye?
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
_______________________________________________________

2. Opaque objects cannot be viewed through the microscope, why?

______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
______________________________________________________________________
_______________________________________________________

Conclusion:
References: