HES 032-Microbiology and Parasitology

College of Nursing
2022-2023
RAD Learning (AY:_________)

Group Leader: MASCARDO, ANNIKA SABRINA

___________________________________

LAZAGA, LOUISE CARATOL

Group Members: _______________________________________
MAGSAYO, JILIAN TRICKSY SULA-SULA
_______________________________________
MAN-ON, ANGELA SWING
_______________________________________
MANGLE, SHANAIA NICOLE LOMA
_______________________________________
_______________________________________

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
th
Keohane, E., Walenga, J., & Smith, L. (2016). Rodak’s Hematology Clinical Principles and Applications. 5 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

Base It helps in holding the various parts of

microscope. It also contains the light source.

C-shaped arm It is used for holding the microscope. And which

is connected the eyepiece to the objective lens.

Mechanical stage It is a rigid platform on which the 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.

B. Illumination and Magnification Parts Function

Illumination Parts:
Used to condense light towards the object.
• Substage condenser Lowering the condenser diminishes illumination
whereas raising the condenser increases the
illumination.

• Iris diaphragm Opening and closing of iris diaphragm controls

the light reaching the object.

It generates a continuous distribution of light

• Light source across the visible spectrum.
Magnification Parts:
It is the lens where the final image of the object is
• Eyepiece (Ocular lens) viewed. Usually, these lenses have a
magnification of either 10X or 15X.

• Objective lens Each objective lens is represented by a particular

3 Types of Objective Lens: 4X (scanning objective)
10X (Low power objective lens).
40X (High power objective lens).
100X (Oil immersion objective lens).
 
colour. 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.

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. 4 x 10 = 40
Scanning = _____________________
b. 10 x 10 = 100
LPO = _________________________
c. 40 x 10 = 400
HPO = _________________________
d. 100 x 10 = 1000
OIO = __________________________

STUDY GUIDE QUESTIONS:

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

The microscope is absolutely essential to the microbiology. Most microorganisms cannot be
seen without the aid of a microscope, save some fungi. It also helps us distinguish disparate
types of bacteria and viruses.

2. Which lens requires the use of cedarwood oil? Why is it important to use oil when
focusing under this lens?
 The objective lens must be designed specifically for oil immersion microscopy. Attempting to use
immersion oil (cedarwood oil) with a "dry" objective will only foul the lens. To use an oil immersion
lens, first focus on the area of specimen to be observed with the high dry (400x) lens. Same
refractive index as glass makes continuous column with slide and objective lens to reduce the loss
of light by refraction and provides more light for 100 x objective lens whose aperture is tiny.
3. What is the difference between bright-phase microscopy and dark-phase microscopy?
Bright-phase microscopy use the most basic and common type of optical microscope. Its
lights source are either LED or halogen lamp. Dark-field or dark phase microscopy is used to
light up unstained samples which can cause it to appear brightly lit against a dark
background. Bright phase microscopy shows a clear magnification while dark phase, on the
other hand shows samples in minute details.
4. Define total magnification and resolution.
Total Magnification of a microscope is the product of the objective and ocular lenses. It is
the ability to make small objects seem larger, such as making a microscopic organism
visible. Resolution is the ability of the microscope to distinguish detail. It is the minimum
distance at which two distinct points of a specimen can still be seen- either by the observer
or the microscope camera.
5. Why do we have to keep both eyes open when using the microscope?
We have to keep both eyes open when using the microscope to avoid fatigue or eye strain,
which occurs when the nonviewing eye is kept closed. It is also important to remember that
the eyes should never touch the ocular lens.

Compiled by: Claire Mae Caval, RMT

Microbiology and Parasitology – Instructor
Southwestern University PHINMA