Name: Aleah A.

Balaoro Course: BSED SCIENCE Date of Submission: 29/09/2022

Laboratory Exercises 3 in Sci Ed 5:

Basic Techniques in Light Microscopy Operation

Rationale:

The microscope is an optical instrument that is used to observe things that are beyond
ordinary vision. It is one of the basic tools in studying biological sciences especially in the field
of microbiology, botany, and zoology.

Objectives:

At the end of the laboratory exercises, it is expected that students can;

1. Identify the mechanical, illuminating, and magnifying parts of the microscope.
2. Identify the function of the different parts of the microscope.
3. Understand the importance of magnification, resolution and illuminations basic
principles of microscopy.
4. Develop skills in the use of microscope
5. Describe the proper care of the microscope

Materials:

Compound light microscope Prepared specimen (plant/animal/microbial cells)

Procedures:

1. Place the microscope close to the edge of the table. Select a suitable stool so that
when looking into the eyepiece, your back is straight, and your neck is bent at the
nape.
2. Lower the body tube by turning the course focus knob until 4x objective (scanning)
reaches the downward stop.
3. Look through the eyepiece and adjust the light source illumination which provides
the optimum brightness, which is the one circular area seen in the eyepiece.
4. Place the specimen on the stage and fasten it using the stage clips.
5. Position the specimen area of the slide over the center of the stage aperture.
6. Looking through the eyepiece, raise the course adjustment knob until the image
appears. Focus as sharply as possible. Scanner or the low power objective has much
greater depth of focus and is generally used for initial focusing and viewing.
7. Adjust the fine adjustment knob to sharpen the image in the center of the field of
vision.
8. When a feature on the specimen is to be examined at a higher magnification, move
the slide so that the feature is centered in the field of vision. The higher the power of
the objective, the lesser is the area of the specimen surface included in the field of
vision. Shift the high-power objective into place and adjust with the fine adjustment
knob.
9. The eye should be kept at certain distance from the eyepiece. Be relaxed when
looking into the eyepiece and keep both eyes open.

V. Self-assessment Questions:
A. Mechanical Parts of Microscope

1. Enumerate the mechanical, magnifying and illuminating parts and its corresponding
functions of the compound microscope.

Answer:

Mechanical Parts

 Base- It helps in holding the different parts of microscope. The light source is also
included.
 Mechanical stage- It is a rigid platform on which specimen to be viewed is placed. It has
an aperture at the center to permit light to reach the object from the bottom. The
object on the slide can be moved either sideways or forward and backward with the
help of the positioning knobs.
 C-shaped arm- It is used for holding the microscope. And which is connected the
eyepiece to the objective lens.

 Diaphragm- The diaphragm is fastened below the stage. It controls and adjusts the
intensity of light that passes into the microscope.

 Body tube- The upper part of the arm of the microscope comprises a hollow and tubular
structure known as the body tube. The body tube can be shifted down and up using the
adjustment knobs.

 Fine adjustment knob- It is the smaller knob, which is used for sharp and fine focusing
of the object. For accurate and sharp focusing, this knob can be used.

  Coarse adjustment knob- It is a large knob that is used for moving the body tube down
and up for bringing the object to be examined under exact focus.

 Nose piece- The nose piece is circular and a rotating metal part that is connected to the
body tube’s lower end. The nose piece has three holes wherein the objective lenses are
embedded.

  Clips- The upper part of the stage is connected to two clips. The slide can be held in its
position with the help of the clips.

  Pillar- It is a vertical projection. This stands by resting on the base and supports the
stage.

 Iris diaphragm- It is seen immediately below the condenser and operated by small
lenses which protrude to one side. Opening and closing of iris diaphragm controls the
light reaching the object.

 Stage- Platform where object to be examine is placed.

Magnifying Part

 Eyepiece or Ocular lens- It is the lens where the final image of the object is viewed.
Usually; these lenses have a magnification of either 4X, 10X or 40X.

 Objective lens: Each objective lens is represented by a color. Here we represent 4X with
red band, 10X with yellow, 40X with blue and 100X with white. These objective lenses
are fitted on to the revolving nose piece. The working distance of an objective is defined
as the distance between the front surface of the lens and the cover glass surface or the
specimen when it is in sharp focus.

-LPO/ Low Power Objective- Gives lowest magnification

-HPO/Higher Power Objectives- Gives higher magnification
-OIO/ Oil Immersion Objective-Gives highest magnification

Illuminating Part

 Substage condenser- It is seen below the stage and made up of a system of convex
lenses which focus light from illuminating sources and is used to condense light towards
the object. Lowering the condenser diminishes illumination whereas raising the
condenser increases the illumination.

 Light source- Light source is situated at the base of the microscope. It is controlled by an
ON /OFF switch and a lamp rheostat. Tungsten-halogen lamps are highly reliable light
source used in the light microscope. It generates a continuous distribution of light across
the visible spectrum.

 Mirror- Located beneath the stage and has concave and plane surface to gather and
direct light in order to illuminate the object.

 Condenser- A set of lenses between the mirror and the stage that concentrates light
rays on the specimen.

2. How does magnifying power of an objective is determined?

Answer:

A common eyepiece lens magnifies 10x, whereas a typical objective lens magnifies
40x. The overall magnifying power of the microscope may be calculated by multiplying
the magnifying powers of the objective lens and the eyepiece, thus 10 x 40 = 400x total
magnification.

3. What is the relationship of magnifying and illuminating power?

Answer:

A two-lens system is used to magnify or enlarge a specimen; the ocular lens is in

the eyepiece, while the objective lens is located in a spinning nosepiece. For effective
magnification and resolving power, effective illumination is important in order to
observe the specimen clearly.
4. Why and how microscopes magnify an object?
Answer:

We use a microscope to magnify or see small objects. It is a scientific tool that

allows you to view details that we cannot perceive with our naked eyes.   When the bent
light reaches the eye, the item seems larger than it is. Objectives are available with
magnifications ranging from 4X–100X.

B. Operation and manipulating the Microscope

5. When you encounter the following problems, what should you do?
Problem Encountered Way(s) to Solve the Problem
a. no image or entirely dark image seen on Make sure you have removed the
the ocular, even though the power protective coverings such as lens covers.
switch is ‘on’ Ensure that the eyepieces are installed. Check
the light or mirror to illuminate the object or
specimen.

b. cannot find the locus/site of the The height of the microscope condenser
specimen that needs to be observed may be set too high or too low. This can also
affect your microscope resolution. You can
adjust or use the Coarse adjustment knob;
this knob is used for moving the body tube
down and up for bringing the object to be
examined under exact focus

c. blurred ocular image of the specimen Clean the microscope's ocular lens with a
being observed lens paper. You can also use Fine adjustment
knob which is used for sharp and fine focusing
of the object. For accurate and sharp
focusing, this knob can be used.
d. double-vision of the specimen being Make sure that the two eyepieces are the
observed correct distance to each other. Adjust the
distance to suit your eyes.
e. bright or highly illuminated image which Adjust the brightness using the brightness
cause teary eyes or fatigue control knob.

C. Proper handling of Microscope

Answer:
 Always carry microscope with one hand on the arm and one arm on the base. Carry it
close to your body.
 Always start and end with lowest powered objectives.
 Do not touch the glass part of the lenses with your fingers. Use only special lens paper
to clean the lenses.
 Rotate the nosepiece of the microscope all the way down to its lowest level when
you have finished using the microscope.
 Store in clean and dry place
 Lower the stage and then remove the slide when you are done. Always clean the
microscope when you are done.
 Cover the microscope with the dust cover to protect it from dirt and other debris.
 Store the microscope on a flat table or surface where it will not be jostled or
knocked over. Ensure that the storage area does not expose the microscope to
prolonged periods of direct sunlight.
 Always place the oculars toward the back of the cabinet.

5. Explain briefly on how to care and handle a microscope?

Answer:

Carry your microscope to your workbench with your two hands, one holding the
arm and one under the base. Carry the microscope and walk, don't run when carrying it.
Check if the bench is clean and dry. Place the microscope down gently toward the center
of the bench to reduce the risk of knocking it off. Ensure that the cord does not hang off
the edge of the bench. Use kleenex or a paper towel to clean any of the surfaces. Use
both eyes to look through the eyepiece of a binocular microscope. Do not touch the
glass part of the lenses with your fingers because this will make them dirty and make
your image blurry. Use only special lens paper to clean the lenses. Plug in and turn on
the microscope. Put the slide on the stage so the specimens are right in the center of
the hole in the stage where the light comes through, then clam the slide in place. Always
follow the low to high rule. Start by selecting the lowest powered objective lens and
leader zoom in by working your way up. Make sure that the specimen is exactly in the
center of the field of view. When your observation is complete, move the turrent to
bring the lower-power objective into position and put the stage down. After using the
microscope, turn it off and unplug it from the power source. Check the stage if it's clean
and dry. Carefully return the microscope to its case and carry it with both hands.

VI. Conclusion: (implication of the over-all knowledge obtained from the activity)
During our exercise, I learnt about the many types of laboratory equipments and
their applications. When doing an experiment, I also learn about the many laboratory
and safety measures.

A microscope is a device that magnifies tiny things. Microscopes are frequently

used in scientific and educational settings to examine items and living creatures that are
not readily visible to the naked eye. It is  important to learn about the different parts of a
microscope to understand how these devices work. We will not be able to get the
intended results if we are unfamiliar with this part and its usage, and the microscope
may be damaged because of improper handling.

This activity helped me to familiarize myself with the microscope's different parts
and functions, as well as its proper use and setup. It helps me gain confidence in using a
microscope, and if I run into problems when using it, at least I have an idea of how to
solve them. 

As future science teachers, it is critical that we understand how to use, handle, and
care for microscopes since we will be able to apply this knowledge in the future.