HES 032-Microbiology and Parasitology

College of Nursing
RAD Learning (AY:23-24)

Group Leader: Amodia, Raul Anthony T.

Group Members:
Agnes, Joanna A.
Aguilar, Faith V.
Aliño, Charls Carlo A.
Alorro, Altheya T.

ACTIVITY NO. 1: The Compound Light Microscope

LEARNING OBJECTIVE: At the end of the laboratory period, the student should be able to:
1. Illustrate the compound light microscope.
2. Recognize the people behind the discovery of the microscope.
3. Identify the basic parts and function of a compound light microscope.
4. Differentiate simple and compound microscope.
5. Compute for the total magnification of the specimen in four different magnifications.

DISCUSSION:

Microorganisms are much too small to be seen with the unaided eye. Hence, they must be
observed with a microscope. The word microscope is derived from the Latin word micro (small)
and the Greek word skopos (to look at). Microorganisms are measured in even smaller units,
such as micrometers and nanometers.The sizes of bacteria and protozoa are usually expressed
in micrometers, whereas the sizes of viruses are expressed in nanometers. A micrometer (mm)
equals 0.000001 m (10^-6 m). A nanometer (nm) equals 0.000000001 m (10^-9 m).

The simple microscope used by Anton Van Leeuwenhoek in the seventeenth century had only
one lens and was similar to a magnifying glass. However, van Leeuwenhoek was the best lens
grinder in the world in his day. His lenses were ground with such precision that a single lens
could magnify a microbe 300x. His simple microscopes enabled him to be the first person to see
bacteria. Contemporaries of van Leeuwenhoek, such as Robert Hooke, built compound
microscopes, which have multiple lenses. In fact, a Dutch spectacle maker, Zaccharias
Janssen, is credited with making the first compound microscope around 1600. However, these
early compound microscopes were of poor quality and could not be used to see bacteria. It was
not until about 1830 that a significantly better microscope was developed by Joseph Jackson
Lister. Various improvements to Lister’s microscope resulted in the development of the modern
compound microscope, the kind used in microbiology laboratories today.

Compiled by: Claire Mae Caval, RMT

Microbiology and Parasitology – Instructor
Southwestern University PHINMA
There are two basic kinds of microscopes depending on the type of electromagnetic wave used
– light microscope and electron microscope. In most laboratories light microscope is a common
instrument. It is further subdivided into two kinds- simple light microscope and compound light
microscope.

A simple microscope is defined as a microscope containing only one magnifying lens. A

magnifying glass could be considered a simple microscope. A compound microscope is a
microscope that contains more than one magnifying lens: objective lens and ocular lens. Light
microscopy refers to the use of any kind of microscope that uses visible light to observe
specimens. Because visible light (from a built-in light bulb) is used as the source of illumination,
the compound microscope is also referred to as a compound light microscope.

According to its function, the basic parts of a compound microscope is divided into two parts:

a. Mechanical parts – comprise all those structures that support the whole device and which
enable the operator to use the illuminating and magnifying parts.
b. Illumination and Magnification parts - consist mainly of lenses (employed as a means of
magnification and glass parts).

Calculate the total magnification of a specimen by multiplying the objective lens magnification
by the ocular lens magnification. Most microscopes used in microbiology have several objective
lenses:
a. Scanning – 4x
b. Low Power Objective (LPO) – 10x
c. High Power Objective (HPO) – 40x
d. Oil Immersion Objective (OIO) – 100x

Most ocular lenses magnify specimens by a factor of 10.

Remember:

Total Magnification = Objective Lens X Ocular Lens

Care of the microscope involves the following details:

1. When not in use for an extended period of time, always cover the microscope to protect it
from dust.
2. Before use, inspect the component parts. If dust is found, use an air syringe, a camel hair
brush, or a soft lint-free cloth to remove it. Using lens paper directly on a dirty lens without first
removing the dust may scratch the lens. Do not use laboratory wipes or facial tissue to clean
the lenses.
3. Avoid placing fingers on the lens surface. Fingerprints affect the contrast and resolution of
the image.
4. Use solvent sparingly. The use of xylene is discouraged, because it contains a carcinogenic
component (benzene). Xylene is also a poor cleaning agent, leaving an oily film on the lens.

Compiled by: Claire Mae Caval, RMT

Microbiology and Parasitology – Instructor
Southwestern University PHINMA
Lens cleaner or 70% isopropyl alcohol employed sparingly on a cotton applicator stick can be
used to clean the objective lenses. Alcohol should be kept away from the periphery of the
lenses, because alcohol can dissolve the cement and seep into the back side of the lens. 5.
When fresh oil is added to residual oil on the 100x objective lens, there may be loss of contrast.
Clean off all residual oil first.
6. Do not use water to clean lenses. If no lens cleaner is available, use a clean microfiber cloth.
7. When transporting the microscope, place one hand under the base as support and one hand
firmly around the arm.
References:

Engelkirk, P., & Engelkirk, J. (2015). Burton's Microbiology for the Health and Sciences. 10th ed., Philadelphia:
Lippincott Williams & Wilkins
Keohane, E., Walenga, J., & Smith, L. (2016). Rodak’s Hematology Clinical Principles and Applications. 5th ed. St.
Louis Missouri: Elsevier Saunders Inc.
Tortora G., F. B. (2016). Microbiology an Introduction, 12th Edition. United States of America: Pearson Education

Compiled by: Claire Mae Caval, RMT

Microbiology and Parasitology – Instructor
Southwestern University PHINMA
OBSERVATIONS:

1. Illustrate and label the basic parts of a compound light microscope.

Compiled by: Claire Mae Caval, RMT
Microbiology and Parasitology – Instructor
Southwestern University PHINMA

2. Tabulate the basic parts of a compound light microscope and their specific functions.
A. Mechanical Parts Function

a. Body tube - holds the eyepiece lens and connects them to

the objective lenses.

b. Revolving nosepiece - holds the different objective lens and facilitates

the changing of objectives.

c. Adjusting knobs - used to focus the microscope.

1. Coarse adjustment knob - moves the body tube and low power lenses
closer or farther away from the stage to view the
image of the specimen. It produces a large
vertical movement of the stage with only a
partial revolution of the knob.

2. Fine adjustment knob - used to bring the specimen into sharp focus to
clearly show the detailed parts of the specimen.

d. Stage - a flat surface where the mounted slide is

placed. A slide is where the specimen is placed
for observation.

e. Stage clips - holds the specimen slide in place.

f. Arm and base - used for support and in carrying the

microscope. Arm allows the body of the
microscope to tilt.

g. Inclination joint - allows the user to tilt the microscope for more
comfortable viewing.

B. Illumination and Magnification Parts Function

Illumination Part

a. Mirror - reflects light from an external light source up

through the bottom of the stage which
illuminates the specimen.

b. Iris diaphragm - controls the amount of light that reaches the

specimen.

- collects and focuses the light from the mirror to

c. Stage condenser the specimen.

Magnification Parts

a. Eyepiece/Ocular lens - viewer looks and sees the magnified image of

the specimen. Have 10x magnification.

- major lenses used for specimen

b. Objective lenses visualization/magnification.

1. Low power objective - has a magnification power of 10x. It is used to

see the general outline of the specimen.

- has a magnification power of 40x, it is used to

2. High power objective
view structures from a larger perspective.

3. Oil immersion objective

- has a magnification power of 100x. It requires
the use of special oil.

Compiled by: Claire Mae Caval, RMT

Microbiology and Parasitology – Instructor
Southwestern University PHINMA
3. Compute for the total magnification using 10x magnification of the eyepiece:

a. Scanning = 4x (objective) X 10x (ocular) = 40x

b. LPO = 10x (objective) X 10x (ocular) = 100x
c. HPO = 40x (objective) X 10x (ocular) = 400x
d. OIO = 100x (objective) X 10x (ocular) = 1000x Microscopy is crucial in microbiology because
it allows scientists to study microorganisms like bacteria, viruses, and fungi at a cellular and
subcellular level. It helps identify, classify, and understand these tiny organisms, aiding
iSTUDY GUIDE QUESTIONS:

1. What is the importance of microscopy to the field of microbiology?

Microscopy is essential to the study of microorganisms in microbiology because it enables

researchers to examine bacteria, viruses, and fungus at the cellular and subcellular level. It
supports illness diagnosis, research, and medical improvements by assisting in the identification,
classification, and understanding of these microscopic creatures.

2. Which lens requires the use of cedarwood oil? Why is it important to use oil when focusing
under this lens?

In a process known as oil immersion microscopy, a high-power objective lens—typically

100x—is required. The numerical aperture and, subsequently, the resolution of the
microscope are improved by using oil. It eliminates light loss and blurring, enabling more
precise and thorough observations..

3. What is the difference between bright-phase microscopy and dark-phase microscopy?

Techniques used in light microscopy include bright-phase and dark-phase microscopy. The
common technique uses bright-phase microscopy, which makes specimens stand out
against a bright backdrop. Contrarily, dark-phase microscopy employs a specific condenser
to produce a black backdrop, allowing transparent specimens to be seen as bright objects..

4. Define total magnification and resolution.

The sum of the magnifications of the objective and eyepiece (ocular) lenses in a microscope
called the total magnification. Resolution is the ability of a microscope to discriminate
between two objects that are separated by a small distance. It is determined by the light's
wavelength and the objective lens's numerical aperture; smaller numbers denote higher
resolution..

5. Why do we have to keep both eyes open when using the microscope?

When using a microscope, keeping both eyes open improves depth perception and lessens
eye strain. In particular when concentrating on three-dimensional specimens, it offers a more
natural and relaxing viewing experience. When using a microscope, spatial awareness is
improved because of how the brain mixes the images from both eyes.
Compiled by: Claire Mae Caval, RMT
Microbiology and Parasitology – Instructor
Southwestern University PHINMA