PRACTICAL 1

TITLE : The Usage of Compound Microscope and The Examination of Eukaryotic Cells.

OBJECTIVES

1. To learn the structure and basic use of the compound microscope.

2. To examine the structure of eukaryotic cells (Plantae and Animalia)

INTRODUCTION

A biologist's essential instrument is the microscope. It was created to aid in examining the
universe of microscopic life forms that are invisible to the naked eye. Early microscopes were
challenging to use and featured only one lens. Magnification was the main issue. The closer
the viewer's eye had to be to the lens, the more potent the lens was required for greater
magnification. The lens was so closely magnified that it almost touched the eye. The first
person using a microscope had to be very steady. The development of the compound
microscope marked a significant advancement in microscope technology. It has two sets of
lenses that magnify things much more than one lens would.

Animal and plant cells differ and are similar in a number of ways. Animal cells, for
instance, lack a cell wall and chloroplasts, but plant cells do. Plant cells have set rectangular
shapes while animal cells are typically spherical and variable in shape. Being eukaryotic cells,
plant and animal cells share a number of characteristics, including the presence of a cell
membrane and cell organelles including the nucleus, mitochondria, and endoplasmic
reticulum. Hypothesis is to examine the structure of eukaryotic cells (Plantae and Animalia)
METHODOLOGY

A. Regular usage and care of compound microscope

1. The microscope was held firmly in both hands, one on the arm and the other
under the microscope's base. Brought to bench in an upright position.
2. The microscope is kept at least 6 inches away from the workbench's edge and two
feet away from any open flame.
3. The dust cover was removed and properly stored.
4. The intensity dial should be set to the least intense position before turning on the
lamp. By doing this, the bulb won't blow. Inform the lab personnel if the
microscope was not operating properly.
5. The position and lighting conditions of the condenser are correct and appropriate.
6. Before and after usage, wipe the lens with a lens cleaning cloth; do not touch any
lenses with fingers.
7. To make sure they are mounted correctly, every objective lens was examined.
Don't divide anything apart.
8. It is guaranteed that every optical component was spotless.
9. Suitable oil must be used for all objective lenses requiring oil immersion.
10. Binocular eyepieces must be used correctly with two eyes, not one, and must be
adjusted to fit the width of the wearer's face. This will produce a single, correctly
aligned image field.
11. Before choosing the targets and during any subsequent transition, the light rays
should be kept away from the eyes.
12. After using the microscope, please ensure that:
a. Lens cleaning tissues were used to clean up the respective objective
lens immediately. A proper lens cleaning solution (e.g.: ether, ethyl
ether, ethanol etc..) may be used when necessary. This step was
particularly important in order to avoid any visible interference, as a
result of debris or oil particles left over during usage, which also
enabled good storage and subsequent usage.
b. The dust covers on the microscopes are properly installed, and they
were stored in the cabinets as directed.
B. Introducing the structure of microscope

1. The Lens System

One of the two sets of lenses is the objective lenses. They work similarly to the lens of
the early, simple microscope. The objective lenses make the initial or primary
magnification. They are located in the nosepiece of the microscope. Inscribed on each
objective is the magnification or power of that lens. This tells the number of times the
lens magnifies the image. For example, if you are looking at a strand of hair with a 4X
lens, the hair will appear four times its actual size. Your microscope probably has at
least two objective lenses. Some microscopes have as many as four objectives. Rotate
the lenses in the nosepiece until they click into position. The objective lens in use is
always the one directly under the body tube.

Usual powers for objective lenses are:

4x The scanning lens

10x The low power lens

40x The high power lens

100x The oil immersion lens

The second kind of lens in the microscope is the ocular – sometimes called the
eyepiece. This lens is located at the top of the body tube. The ocular serves as a small
telescope, magnifying the image made by the objective lens. This enlargement is
called the secondary magnification. The magnification of the ocular may be 5X, 10X,
15X, or 20X. The most common power used in microscopes is the 10X ocular.
Examine the ocular of your microscope. Do not remove it from the body tube. If the
power is not stamped on the top portion of the ocular, you should assume that it is
10X. The total magnification of the microscope is determined by multiplying the
primary magnification (from the objective) by the secondary magnification (from the
ocular).
 2. The Stage

A specimen to be viewed through the microscope is mounted on a glass slide and

covered with a cover slip. The slide rests on the stage, the flat surface beneath the body
tube. Stage clips hold the slide in place. Also, they help in making slight adjustments in
the slide's position by holding the slide steady. The stage should always be kept in a
horizontal position. If you tilt the stage, the specimen will slip to the bottom edge of
the slide. Even with commercially prepared slides, the stage should be kept horizontal.
A commercial slide can be ruined as the cover slip slowly slips downward on a tilted
stage. Both the slide and the stage are extremely smooth. Water between them acts like
glue and causes the slide to stick to the stage. If water gets on the stage, STOP, and dry
both the stage and the bottom of the slide with a paper towel before proceeding.

3. The Lighting System

For you to see the specimen, light must pass through it and the lenses to your eye. The
lighting system is located under the stage of the microscope. There are three different
types of lighting systems. The simplest system uses a concave mirror to focus a beam
of light on the slide. Tilt the curved surface of the mirror to face a light source: room
lights, windows, or a desk lamp. Another lighting system uses a lens under the stage to
focus the light. If there is a substage lens and a mirror on your microscope, use the flat
side of the mirror to reflect light through this lens. A third lighting system uses a
substage light instead of a mirror. If your microscope has a light, turn it on only when
you are actually looking at the specimen. The light gets hot and can easily destroy
your specimen.

Under the stage you will also find the diaphragm. It is used to adjust the amount of light
that passes through the specimen. The diaphragm works like the aperture on a camera.
Practice opening and closing the diaphragm while looking through the eyepiece. Notice
how the amount of light increases and decreases.
 4. The Focusing System

In order to bring the image of the specimen into proper focus, it is necessary to change
the distance between the slide and the objective lens. This can be done in one of two
ways, depending upon the microscope you are using. Either the lenses can be moved or
the stage upon which the slide rests can be moved. Two knobs control the focus.The
coarse adjustment knob is for coarse focusing, and the fine adjustment knob is for fine
focusing. Locate these on your microscope. Turn the coarse adjustment knob.

C. Using the compound microscope

1. Put the low power objective in place. Look through the ocular and adjust the light so
that you see a uniformly bright field of view. The field of view, also called the field, is
the area you see through the lens. If you see specks of dirt in the field, clean your lenses
with lens tissue.
2. For microscope viewing, slides were made. Cut up from an old newspaper, the
lowercase "e" was positioned in the middle of a clear glass slide.
3. On the letter, a drop of water was applied.
4. The cover slip is then lowered slowly over "e" after having its edge put on the water.
Cover slip was positioned in this manner to stop bubbles from forming.
5. Be sure that the bottom of the slide was dried. This type of slide was called a wet mount.
6. The slide was positioned beneath the stage clips such that the "e" was upright.Able to
concentrate on the "e."
7. With the lesser power (4X and 10X) objectives, focusing always starts. The low power
goal was first placed in the nosepiece and snapped into place. When the objective was as
close to the slide as feasible without touching it, the coarse adjustment knob was moved
while keeping an eye on the side of the microscope.
8. The coarse adjustment knob was moved while looking through the ocular to shift the
objective farther from the stage. The approximate shape of the "e" became apparent. The
fine adjustment knob was rotated back and forth to sharpen the focus.
9. Through the microscope, take note of where the letter is located. The letter was upright
on the slide.
 10. The "e" was now examined in fine detail. First, centre the "e" in the area of view while
using low power. By rotating the nosepiece until the high power objective clicks into
position, you could change it to high power.
11. By adjusting the fine adjustment knob, the focus was made more precise.
12. Try this if you're having trouble finding the "e" under high power. The slide was slightly
moved while the ocular was looked through. If doing so does not reveal the "e," the
slide shifts in a different direction.
13. Observation was recorded by both low and high power objective lenses.
14. The whole procedures were repeated with the prepared slides of plant cells and animal
cells.

DATA AND ANALYSIS

Observation

The letter ‘e’

Power of Objective Lens 40x

Eukaryotic Cells Plantae Animalia

Observation

Name of Eukaryotic Cells Monocot Docsend Stem C.S Mil. Tubercul (Liver)

Power of Objective Lens 40x 40x

CONCLUSION

Based on the result,we use two types of eukaryotic cells,plantae and animalia. For plantae, we
use Monocot Drocdend Stem C.S while for animalia,we use Mil Tubercul (Liver). Power of
objective lenses we use are 40x. We use that power because it is easier for us to observe
eukaryotic cells by using a microscope. We can use another power of objective lenses but we
use 40x because we want to observe fine details within a specimen sample. If we use low
power of objective lenses, for example 10x objective lens. One of the most useful lenses for
monitoring and evaluating glass slide samples is the low power objective lens, which has more
magnification power than the scanning objective lens. We don’t use the highest objective lens
because the high-powered lens can become damaged as it’s very close to the slide.

In this experiment, the hypothesis is accepted.

QUESTION

1. Do the lenses move up and down or does the stage move up and down?

The stage moves up and down.

2. Is the position of the letter viewed through the microscope the same as it is on the stage?

The position of the letter ‘e’ viewed through the microscope is the same as it is on the
stage.

3. What is the relationship between movement on the stage and movement seen through
the lenses?

The movement on the stage is diametrically opposed to the movement seen through the
lenses. For instance, if you move something on the left on the stage, what you see
through the lenses is the ‘specimen’ moving to the right.

4. Note the length of each lens. Is the higher power lens longer or shorter than the lower
power lens?

On low power, you will see more of an object. The lowest power objective has the
greatest depth of focus. When you increase the power, the depth of focus decreases. As
a result, at higher magnification, a smaller portion of the specimen is in focus.