LABORATORY ACTIVITY NO.

2
MICROSCOPY

Group Leader: Magcalas,Carlos Jose B. Date Performed: 08/23/2021

Members: Javier, Romelyn Joyce Date Submitted: 08/31/2021

Llanto, Anne Benadeth
Monterozo, Audrey Rose
Olave, Kyla Denise

The microscope is a very powerful and useful tool in microbiology. However, to

maximize its use, it is necessary that a proper knowledge and skill must be acquired.

OBJECTIVES

At the end of this laboratory activity, the students will be able to:

1. review the parts of the microscope;

2. apply the important terms used in microscopy;
3. differentiate the properties of various microscope types;
4. calibrate ocular micrometer divisions;
5. compute the sizes of some representative microorganisms.

I. Parts of the Microscope

Label the different parts of the microscope, and fill up the table below.
6.
A. Parts of the Microscope

7. Ocular Lens
7
(Eyepiece)
1Body Tube

2 Revolving Nosepiece
2. 8. Arm
8
3
3. Objectives
9. Stage
9
4. Stage Clips
4
10. Coarse
10 Adjustment Knob
5 Condenser
5.
11. Fine11
Adjustment Knob
6 Source
6. Light

12Base
12.
 B. Functions of the Microscope
Parts of the Microscope Function
1. Body Tube Connects the eyepiece to the objective lenses.
2. Revolving Nosepiece The part of the microscope that holds two or more objective lenses that
can be rotated to easily change power
3. Objective lenses These are the major lenses used for specimen visualization

4. Stage Clips Holds the specimen slides in place

5. Condenser Lenses that are used to collect and focus light from the light source into
the specimen.
6. Light Source Shines up through the slide

7. Ocular Lense The top of the microscope where the viewer looks through

8. Arm Connects the eyepiece tube to the base

9. Stage Flat platform that supports the slides

10. Coarse Adjustment Allows quick focusing by moving the objective lens or stage up and
Knob down

11. Fine Adjustment Knob Used to fine focus the image when viewing at the higher magnifications
12. Base Acts as microscope’s support

II. Terminologies in Microscopy

Answer the following questions.
1. A microscope has an eyepiece lens with a power of 10X. The lower objective lens being
used has a power of 4X.
Total magnification = 40X
2. A microscope has an eyepiece lens with a power of 16X. The medium objective lens
being used has a power of 10X
Total magnification = 160X
3. A microscope has an eyepiece lens with a power of 12X. The higher objective lens being
used has a power of 40X.
Total magnification = 480X
4. Always begin examining microscope slides with which power objective? Justify your
answer.

a) high; b) low; c) 100x; d) It doesn’t matter which objective

Answer: When examining your specimen under the microscope, you must always begin observing
them using the low power objective. Starting with this objective would provide you a wider field
of view, this would help you locate your specimen a lot easier. After locating the structure that is
needed to be observed and putting it at the center, you finally can switch to the higher objectives
for higher magnification.

5. You are planning to create a better compound light microscope—one that will enable
you to see objects smaller in diameter than 0.2 µm. You gather together the best lens
grinders in the world and put them to work in a lens-grinding laboratory having
unlimited resources. You instruct them to grind marvelous magnifying lenses and add
them to an existing compound light microscope. What’s wrong with this plan?
Answer: There are two things that are wrong in this plan. First, considering that the compound
microscopes use light to produce images and visible light has certain wavelengths (380nm -
750nm), it limits the microscope to produce images that are smaller than the half of the length
of the light waves. This means that it is impossible to make a compound light microscope that
enables someone to see objects smaller than 0.2 µm even using marvelous magnifying lenses.
Second, adding magnifying lenses to an existing compound light microscope is not also a good
plan. Given the fact that the compound light microscopes have limitations that were stated
previously and you have unlimited resources, you might as well create a different kind of
microscope that does not use visible light to produce images of objects smaller than 0.2 µm.

6. While using a compound light microscope, you have noticed that additional light is
required as you increase magnification. Describe three correct ways to increase the
amount of light entering the objective lens, and one incorrect way.

Answer: Increasing or reducing the light intensity (using the microscope rheostat control, moving
the condenser lens closer to or away from the sample (using the condenser adjustment knob),
and opening/closing the iris diaphragm are the three main ways to change the brightness. When
one has a condenser adjustment knob, it is incorrect to adjust the knob while using the
microscope to avoid the sample being in the wrong position.

III. Types of Microscopes based on Properties and Functions

Fill in the table with the needed information:

Characteristics of the various types of microscopes.

Type Resolving Typical Uses of Microscope Sample Image Taken under the
Power Microscope (write the type of
organism)
Bright-field 0.2 μm ● used for viewing live and
stained specimen

Tooth enamel formation of an

immature pi
Dark-field 0.61 I/N.A. ● used for demonstrating of
very thin bacteria that is
not visible under ordinary
illumination and of the
motility of flagellated
bacteria and protozoa

Marine bryozoan Pectinatella

Phase 0.2 μm ● used to observe live and
Contrast unstained specimens
● can view internal cell parts
such as mitochondria,
lysosome, and the Golgi
body

Whipworm eggs (Trichuris

trichiura)
Differential ● 0.24 μm ● used for rendering
Interference contrast in transparent
Contrast samples and gives brilliant
pseudo-3D relief shading
images

Whipworm eggs (Trichuris

trichiura)
Fluorescent ● about ● used in fading, quenching,
180 nm and photobleaching
in focal ● used in fluorescent-
plane antibody technique which
● about can identify unknown
500 nm bacteria
along ● used to visually enhance
the 3-D features at small
optical scales such as microbes Mouse kidney tissue
axis
Confocal 0.5 to 1.5 μm ● has the ability to collect
consecutive optical
sections from a thick
specimen
● used in image degradation
of the specimen

Dendritic cells
Transmission 1 nm ● used to observe the
Electron detailed internal
Microscope structures of cells

Several peripheral myelinated

fibers and a Schwann cell
Scanning 1nm ● used in observing
Electron specimens in three
Microscope dimensional images

Radiolarians
IV. Calibration of the Ocular Micrometer on a Microscope

Determine the calibration factors of each of the microscopes described below. Show solutions
(you can solve on a piece of paper, and then attach the photo of your solution to the google
classroom assignment) and write here the final answer.

1. At a total magnification of 40x, a student measured 42 ocular micrometer divisions per

millimeter. What is the distance in micrometers per ocular unit?

Answer: 23.81 μm per ocular unit

2. At a magnification of 40x, a student measured 41 ocular micrometer divisions per

millimeter. What is the distance, in micrometers, per ocular unit at 40x?

Answer: 24.39 μm per ocular unit

3. At a total magnification of 100x, a student measures 16.4 ocular micrometer divisions per
millimeter. What is the distance, in micrometers, per ocular unit at 100x?

Answer: 60.98 μm per ocular unit

4. At a total magnification of 400x, a student measures 4.1 ocular micrometer divisions per
millimeter. What is the distance, in micrometers, per ocular unit at 400x?

Answer: 24.39 μm per ocular unit

 5. It was found that the units on the ocular micrometer and the units on the stage
micrometer matched up 40 units to 1.0 mm at 100 power. (So 40 ocular units equals 1.0
mm.) If the wing of a dead fruit fly was measured to be 2.5 ocular micrometer units at a
magnification of 100, what is the length of the wing in mm? What does mm stand for?

Answer: The wing is 0.0625 mm in length, mm stands for millimeters

6. Upon computing all the values for ocular divisions using all the objective lenses that came
with your microscope, is it necessary to always re-calibrate the lens every time you use
the microscope? When will it be necessary?

Answer: Every magnification has its own calibration settings, re-calibration is necessary when
the total magnification is changed. If images or measurements are taken using incorrect
calibration settings, the magnification indication and measurement values will also be incorrect,
requiring the whole process to be performed again.

V. Measurement of Microscopic Samples

Presented in the table below are the calibration factors presented in the video,
“Calibration of the Microscopic Ocular Micrometer” by Gilles Buldoc in 2017.

Calibration Factor of the as presented by Gilles Buldoc, 2017:

Ocular lens Objective lens Total Calibration

magnification magnification Magnification Factor, um
10x 4x 40x 25
10x 10x 100x 10
10x 40x 400x 2.5

Determine the sizes of the following samples viewed under the microscope:

1. You are viewing the red blood cells under the 400x magnification. What is the
diameter/length of the red blood cells that are being pointed by the arrows?

1a

1b
Answer: a= 20 μm in diameter/length
b= 27.5 μm in diameter/length
Show how you computed this. You can do the computation on a paper, and you may attach a
photo to the google classroom of how you solved this.

Answer:
2. You are viewing the following cell in the scanner. What is the length of the sample?

Answer: The sample is 250 μm in length

Show how you computed this. You can do the computation on a paper, and you may attach a
photo to the google classroom of how you solved this.
 3. You are viewing the following cell in the high-power objective. What is the length of the
sample?

Answer: The sample is 185 μm in length under high power objective

Show how you computed this. You can do the computation on a paper, and you may attach a
photo to the google classroom of how you solved this.
Conclusion:

Microscope is an essential equipment inside the microbiology laboratory thus, it is

significant to learn all about this equipment. After answering the given tasks in each part, the
purpose of the activity has been met. Different parts of the microscope have been reviewed using
the picture given as well as their specific functions. Terms in microscopy were answered which
gave way in gaining knowledge about total magnifications and specific solutions to analyzation
problems, specifically the proper use of the microscope. Types of microscopes were also learned
by answering a table which contains characteristics for each microscope type. Calibration factors
were also determined and sizes of some representative organisms were computed.