Name: _______________________________________ Date: _______________________

Exercise 1 Light Compound Microscope

The compound light microscope is an important tool in the study of cells, tissues and organs. It is designed
to enlarge the image of these structures that may be too small to be seen by the naked eye. It is important to know
the parts and functions of the microscope, to understand its proper use and care. There are several designs of the
compound microscope, but all of them consist of magnifying (lens system), illuminating (illumination system)
and mechanical (geared focusing system) parts.

Figure 1 shows
the main parts of
two types of
commonly used
compound
microscopes.

The Lens
System

The
compound
microscope has a
lens system, described as magnifying parts, which function to enlarge objects. It consists of two kinds of lenses:
the ocular lens or eyepiece located at the upper end of the body tube and the objective lenses located just below
the body tube. The ocular lens, inscribed with a 10x at its top side, magnifies the image of the specimen by 10
times. Generally, there are two objective lenses attached to the revolving nosepiece that allows them to be rotated
in or out of position with reference to the slide on the stage. When rotating the revolving nosepiece, look from
the side and make sure that the lenses do not hit the stage. The shorter objective, referred to as the low power
objective (LPO), with an inscribed 10x at its side, has a magnification of 10X. The longer objective lens, referred
to as the high power objective (HPO) may be inscribed with a 40x or 60x at its side which also indicates its
magnification. If the microscope has 3 or more objectives, these may include a scanner with a magnifying power
of 4x and/or an oil immersion lens with a magnifying power of up to 100x.

The Illumination System

The illumination system consists of the following:
a) An adjustable mirror that reflects the light from a source into the condenser.
b) The condenser beneath the stage that concentrates light to provide proper illumination to the field of view.
The condenser may be raised or lowered using the substage knob.
c) The iris diaphragm is a thin structure with an adjustable aperture located at the mouth of the condenser.
It regulates the amount of light that enters the condenser. The size of the aperture is regulated by the iris
diaphragm lever located at the side of the condenser
Mechanical Parts
Mechanical parts function to support, hold or help in the adjustment of parts of the microscope.
The drawtube holds the eyepiece. The body tube provides appropriate distance and ensures proper alignment
between the eyepiece and the objectives. The revolving nosepiece supports the objectives and facilitates shifting
from one objective to another to achieve desired magnification of the object under observation. The dust shield
is set above the revolving nosepiece and provides a protective covering for unused objectives, preventing dust
from settling and collecting on their surfaces.
The stage is the platform where the mounted specimen is placed for observation. It is generally provided with
a pair of clips to hold the slide in place. In some models, a mechanical stage may take the place of the clips.
The substage supports the condenser and the mirror. It may have a substage knob that can be used to adjust
the position of the condenser between the stage and the mirror.
The arm connects the body tube to the pillar. The pillar is connected to the base and provides major support
for the microscope. An inclination joint may be found between the arm and the pillar, and functions to allow the
tilting of the microscope for comfortable viewing. When transporting a microscope from one place to another,
make sure to hold the arm with one hand and support the base with the other hand.
Proper focusing requires the attainment of the correct distance between the objective lens and the slide. This
distance is called the working distance. Adjusting the distance between the objective lens and the slide is carried
out by means of the geared focusing mechanism consisting of two rounded knobs located on each side of the
microscope. The larger knob, referred to as the coarse adjustment knob, is used for raising or lowering the
objective lens to a considerable distance. The smaller knob, referred to as the fine adjustment knob, is used for
critical focusing. Note that turning the right knobs clockwise will shorten the distance between the objective lens
and the stage. On the other hand, turning the right knobs in a counter clockwise direction will move the lens away
from the stage. In some models, it is the stage that moves up or down with the objective lens remaining stationary.
Proper use of the microscope
a) Place the microscope in front of you, no less than an inch and no more than two inches from the edge of
the table.
b) Turn the nosepiece so that the LPO is in line with the tube; then lower the tube as far as it will go.
c) Adjust the iris diaphragm to give maximum opening.
d) While, looking through the eyepiece, adjust the mirror until the field of view is properly lit. Proper
illumination means the field of view has uniform lighting. If the microscope has a condenser, the plane
mirror should be used with the HPO; the concave mirror may prove more satisfactory with the LPO,
especially when a desk lamp is used as a light source. If the microscope does not have a condenser, the
concave mirror should be used with all objectives.
e) Place the prepared slide on the stage. While looking through the eyepiece, move the slide until the desired
part of the specimen is positioned in the center of the circular field of view. Use the stage clips to keep the
slide in place. Slowly turn the right coarse adjustment knob clockwise until the image comes into focus.
For sharp and clear image, use the fine adjustment knob to get the proper working distance.
f) For viewing under higher magnification, use the revolving nosepiece to shift from LPO to HPO. Note that
the HPO is longer than the LPO. To prevent damage to the microscope or breakage of prepared slides,
look from the side and make sure the HPO fits nicely into the space above the stage.
g) Most well-maintained microscopes are parfocal, which means that when the object is in clear focus under
LPO, it will also be in clear focus under HPO. Thus shifting from one objective to another will not require
any other focusing adjustment.
h) If the microscope is not parfocal, make sure to lower the HPO very carefully until it touches the upper
surface of the mounted specimen. Then, using the adjustment knob, very slowly increase the distance
between the HPO and the slide until the specimen comes into clear focus.
Care of the Microscope
a) Examine the microscope and report any damage immediately.
b) When transporting the microscope, it should be carried in an upright position with one hand holding the
arm and the other hand supporting the base.
c) Before putting away the microscope, adjust the LPO so that it is in line with the body tube and lower it
until it is as close to the stage as possible. Adjust the stage clips parallel to each other, close the iris
diaphragm and put the mirror in a vertical position.
d) The eyepiece should not be removed from the tube. If the unexposed lens of the eyepiece requires cleaning,
inform the instructor.
e) Lens are cleaned only with lens paper.
 f) If any liquid is spilled on the stage, or comes in contact with the lenses, wipe it off immediately. Make
certain that any liquid that may have dripped down below the stage is also removed.
g) The microscope is an expensive precision instrument, it is your responsibility to use it properly.

ACTIVITY SHEET 1
I. Label the parts of the microscope

II. Briefly answer the following questions.

Exercise 2 Preparation of Temporary or Wet Mounts

The main purpose of this exercise is to enable students to prepare specimens for microscopic examination.

MATERIALS: beaker with water, medicine dropper, glass slides, cover slips,

PROCEDURE:

1. Place a small drop of water on a clean slide.

2. Place the specimen in the drop of water.
3. Cover the specimen with a cover slip by first touching the slide with one edge of the cover slip and then
lowering the other edge. This prevents the formation of bubbles.
4. Water should fill the space between the slide and the cover slip; if it spreads over any other part of the slide,
this excess should be blotted carefully.
5. If the material is already in a liquid suspension, a drop of the suspension can be used without step number 1.
6. It is often desirable to replace the water in which the specimen is mounted with some other medium without
disturbing the mount. Add a drop of the new medium to one edge of the cover slip and draw it under the cover
slip by laying blotting paper along the opposite edge of the cover slip.

Exercise 3 Magnification

The specimen is enlarged by a lens or a lens system of the microscope. In this exercise, students will be able
to estimate the number of times an object viewed under the microscope is enlarged or reduced in size in a drawing.

Magnification of the Image of an Object

The image of an object observed under a compound microscope is enlarged by both the eyepiece and the
objective lens. The magnification of a microscopic object is equal to the number of times it is magnified by the
eyepiece multiplied by the number of times it is magnified by the objective lens. This kind of magnification is
known as linear magnification.

Magnification of a Drawing

Magnification of a drawing is determined by using the following equation:

Magnification = size of drawing

size of specimen
where the size of the drawing and the size of the specimen are expressed using the same unit of measure.
Microscopic objects are measured with a micrometer eyepiece where the unit of measurement is in micrometer
spaces (msp). The magnification of the drawing of a microscopic object may be determined by using the following
equation:

Under LPO
Magnification = size of drawing in millimeters
size of specimen in micrometer spaces x 0.015 mm/msp

Under HPO
Magnification = size of drawing in millimeters____________
size of specimen in micrometer spaces x 0.0036mm/msp

Drawings in the laboratory do not require a talent for illustrating. They are made only as a means to enable
the student to grasp and show basic ideas simply and clearly. Even an unsteady hand can make informative
drawings if the following rules are observed:
1. Begin you drawing with some light lines to bring the parts into their proper relationships. For example, if the
object is roughly rectangular, draw a rectangle then outline the main parts of the object.
2. You are now able to make the final outlines by referring back to the specimen.
a. Use the eraser freely to remove the rough lines.
b. Draw with a sharp pencil to make clean lines.
c. It should not be necessary to shade.
3. Label your drawings fully.
a. Use a ruler to make straight pointer-lines that end on the object designated.
b. Print your labels so that they are legible.
c. Make your pointer-lines parallel, and try to make them end on the same vertical line.

MATERIALS: compound microscope, glass slide, cover slip, tap water, micrometer eyepiece and ruler.

SPECIMENS: Santan leaf, Rice grain and pollen grains from Gumamela

PROCEDURE:

A. Macroscopic Objects
1. Measure the length and width of the leaf in millimeters. Draw the leaf 40 mm in length. Compute for the
proportional width before making your drawing. Determine the magnification of your drawing.
2. Repeat procedure A1 for the rice grain, but make the length of the drawing 20 mm. Indicate the magnification
of your drawing.

B. Microscopic Objects
1. Make a wet mount of the pollen grains of gumamela. Examine and measure the diameter of a pollen grain
under the LPO using a micrometer eyepiece. Draw the pollen grain 20 mm in diameter. Compute for the
magnification of your drawing.
2. Repeat the above procedure but this time use the HPO and make your drawing 40 mm in diameter

OBSERVATION AND DISCUSSION:

I. Drawings

___________
II. Answer the following questions.

1. Describe the direction of movement of the image of the pollen grain when you move the slide:
a. to the right ______________________________________________________
b. to the left _______________________________________________________
c. towards you _____________________________________________________
d. away from you ___________________________________________________
2. What happens when the size of the aperture or opening of the iris diaphragm is:
a. reduced _________________________________________________________
b. increased ________________________________________________________
3. Which objective lens covers a wider field? _________________________________
4. Which objective lens gives a larger image? _________________________________