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Microbiology

Rosie Lynn P. Tejada

Institute of Aquatic and Applied Sciences
Course Title Microbiology

Microbiology Course Pack 1

Course Descriptions This course deals on the study of bacteria, yeast,
molds, microscopic algae and microscopic parasites
associated with fish: their characteristics. Likewise,
it would deal on the principles and practice of
important microbiological techniques.

Units/Credit 3 units (2 units –Lecture, 1-unit Laboratory)

Equivalent

Course Outcomes The students will be able to learn the major concepts
and principles of microbiology. It will also enable the
students to infer the importance of the learned
theories to various applications.

Learning Outcomes At the end of the course, the students will be able to:
 Articulate on the different concepts of
microbiology;
 Categorize the different microorganisms and
identify the different microscopic parasites in
fish;
 demonstrate the different microtechniques;
 realize the importance of microorganisms and the
practical effects of their activities to the higher
forms of animals.

Microbiology Course Pack 2

 TABLE OF CONTENTS

Lessons Page

Module 1: Introduction to Microbiology and Microscopic 4

Methods
Module Overview 4

Lesson 1. Definition and Importance of Microbiology 5

Lesson 2. Scope and Fields of Microbiology 8

Lesson 3. Microscopic Methods 15

Module Summary 25

References 26

Microbiology Course Pack 3

 Module

1
Introduction to Microbiology and Microscopic
Methods

Module Overview

Welcome to the introductory lesson of Microbiology. This module covers the

information about the general concepts and principles of Microbiology.
Microbiology is involved in various fields such as Pharmacy, Medicine, clinical
research, agriculture, dairy industry, water industry, nanotechnology and chemical
technology.
Microbiology is the study of microscopic organisms either single-celled (unicellular),
cell colony (multicellular) or acellular (lacking cells). Microbiology includes many sub-
disciplines like virology, mycology, parasitology and bacteriology.
After the completion of this module, you should be able to:
1. Define Microbiology
2. Discuss the importance of Microbiology
3. Discuss the Scope of Microbiology.
4. Enumerate the fields of microbiology.
5. Identify the parts of the microscopes; relate how to care the microscope and
enumerate the types of microscopes.

Module 1 composed of the following Lessons:

Lesson 1: Definition and importance of Microbiology.

Lesson 2: Scope, Fields and History of Microbiology
Lesson 3: Microscopic Methods

Microbiology Course Pack 4

 Lesson 1. Definition and Importance of Microbiology

Learning Outcomes

At the end of the lesson, you should be able to:

1. Define Microbiology.
2. Discuss the importance of Microbiology.

Time Frame: 1 week (5 hours in a week)

Introduction

Welcome to Lesson 1!
This lesson orients you on the meaning of Microbiology.
Microbiology is the study of all living organisms that are too small to be seen with the
naked eye. This includes bacteria, archaea, viruses, fungi, prions, protozoa and algae.
They are collectively known as 'microbes'.

Activity

What do you know already?

1. What is your basic knowledge about microorganisms?
Open the link below, watch the video and answer the questions in the analysis part.
https://www.youtube.com/watch?v=fU0XO1X1tAE
https://www.youtube.com/watch?v=qYDQ8JjEeF0

Analysis

1. What are the differences between prokaryotes and Eukaryotes?

2. What are the most significant information you learned in watching
and listening the 2 videos?

Microbiology Course Pack 5

 Abstraction

Please read carefully!

Micro-organisms and their activities are really important to almost all processes on
Earth. Micro-organisms affect every aspect of our lives – they are in us and around us.

Microbiology is the study of microorganisms, or microbes, a diverse group of generally

minute simple life-forms that include bacteria, archaea, algae, fungi, protozoa,
and viruses. The field is concerned with the structure, function, and classification of
such organisms and with ways of both exploiting and controlling their activities.

These microbes play key roles in nutrient cycling, biodegradation/biodeterioration,

climate change, food spoilage, the cause and control of disease, and biotechnology.
Thanks to their versatility, microbes can be put to work in many ways: making life-
saving drugs, the manufacture of biofuels, cleaning up pollution, and
producing/processing food and drink.

Microbiologists study microbes, and some of the most important discoveries that have
strengthened modern society have resulted from the research of famous
microbiologists, such as Jenner and his vaccine against smallpox, Fleming and the
discovery of penicillin, Marshall and the identification of the link between Helicobacter
pylori infection and stomach ulcers, and zur Hausen, who identified the link between
papilloma virus and cervical cancer.

Microbiology research has been, and continues to be, central to meeting many of the
current global aspirations and challenges, such as maintaining food, water and energy
security for a healthy population on a habitable earth.

Microorganisms are extremely important in our everyday lives. Some are responsible
for a significant proportion of the diseases affecting not only humans, but also plants
and animals, while others are vitally important in the maintenance and modification of
our environment. Still others play an essential role in industry, where their unique
properties have been harnessed in the production of food, beverages and antibiotics.
Scientists also have learned how to exploit microorganisms in the field of molecular
biology, which makes an enormous impact both industrially and medically.
Microbiology also encompasses immunology, the study of the body’s ability to mount
defenses against infectious microbes.

Importance of Microbiology
 Microorganisms are indispensable components of our ecosystem. Microorganisms
play an important role in the recycling of organic and inorganic material through
their roles in the C, N and S cycles, thus playing an important part in the maintenance
of the stability of the biosphere.
 They are also the source of nutrients at the base of all ectotropical food chains and
webs. In many ways all other forms of life depend on the microorganisms.

Microbiology Course Pack 6

 Microorganisms also have harmed humans and disrupted societies over the
millennia. Microbial diseases undoubtedly played a major role in historical events
such as decline of the Roman empire and conquest of the new world.
 In addition to health threat from some microorganisms many microbes spoil food
and deteriorate materials like iron pipes, glass lenses, computer chips, jet fuel, paints,
concrete, metal, plastic, paper and wood pilings.

SUBDISCIPLINES OF MICROBIOLOGY

Bacteriology
This is the study of bacteria.

Environmental Microbiology
This is the study of the function and diversity of microbes in their natural environments.

Evolutionary Microbiology
This is the study of the evolution of microbes.

Food Microbiology
This is the study of microorganisms causing food spoilage as well as those involved in
creating foods such as cheese and beer.

Industrial Microbiology
This is the exploitation of microbes for use in industrial processes, such as industrial
fermentation and wastewater treatment. This is linked closely to the biotechnology
industry.

Medical (or Clinical) Microbiology

This is the study of the role of microbes in human illness. It includes the study of
microbial pathogenesis and epidemiology and is related to the study of disease
pathology and immunology.

Microbial Genetics
This is the study of how genes are organized and regulated in microbes in relation to
their cellular functions. This subdiscipline is related closely to the field of molecular
biology.

Microbial Physiology
This is the study of how the microbial cell functions biochemically. It includes the study
of microbial growth, microbial metabolism and microbial cell structure.

Mycology
This is the study of fungi.

Veterinary Microbiology
This is the study of the role in microbes in veterinary medicine.

Microbiology Course Pack 7

Virology
This is the study of viruses.

Application

1. Is Microbiology important to other sciences? Discuss briefly.

2. Can you cite advantages and disadvantages of studying

microorganisms.

Congratulations! You did a great Job! You have finished the activities and tasks
for Lesson 1. It is expected that you have gained additional information about
Microbiology . Rest and relax for a while then move on to Lesson 2. Scope and Fields
of Microbiology. Good Luck!

Lesson 2. Scope and Fields of Microbiology

Learning Outcomes

At the end of the Lesson, you should be able to:

1. Discuss the scope of Microbiology.
2. Relate the importance of the different fields of Microbiology.
3. Describe the two main Branches of Microbiology.
Time Frame: 1 hour

Introduction

Welcome to Lesson 2.
In this lesson, you will learn the scope and fields of Microbiology.
Microorganisms are important to all of us in a multitude of ways. The impact of
microorganism in human life and in the environment is both beneficial as well as
detrimental also.

Microbiology Course Pack 8

 Activity

Please open the link below, watch and listen the video and answer the question in the analysis
part.

https://www.slideshare.net/SnehalPatel98/introduction-to-microbiology-84925450

Analysis

What did you learn in the video you are watching and listening?

Abstraction

Please read carefully!

Fields or branches:

Microbiology has two major fields

1. Pure Microbiology
2. Applied Microbiology

1. Pure microbiology: (on the basis of Taxonomical classification)

 Bacteriology
 Mycology
 Phycology
 Virology
 Protozoology
 Immunology
 Parasitology

Bacteriology is a branch of Microbiology that deals with study of Bacteria. This

Bacteria are prokaryotic, unicellular in nature. Their mode of multiplication is by
Binary fission. Bacteria can be parasitic or can be free living in atmosphere. The nuclear
material is not bound to nuclear membrane. This bacteria may be motile or non-motile.
They may vary in their shape like rod, spiral or cocci and they may be aerobic, non-
aerobic or facultative anaerobic in nature. On the basis of mode of nutrition some
bacteria may have autotrophic or hetrotrophic mode of nutrition.

Microbiology Course Pack 9

Mycology is a branch of microbiology that deals with study of Fungus. This fungus
cells are eukaryotic in nature the nuclear material is surrounded by Chitin or cellulose
or both. This fungal cells are non-photosynthetic and chemoorganotrophic in nature.
These fungal cells are divided into two types and that is yeast and molds.

 Yeast -The yeast cell may occur in single cell or pseudomycelium form. The
mode of reproduction is by budding or by Spore formation. Yeast are also know
as Ascomycetes and this yeast cells may be oval, rod or spherical in shape.
 Molds-The molds grow in form of multi-cellular filamentous structure called as
hyphae. This hyphae may be septed or non-septed. They can reproduce by both
means of sexual and asexual mode of reproduction.

Phycology is a branch of microbiology that deals with the study of algae. They are
photosynthetic, eukaryotic, and multi cellular organism.

Virology. This branch of microbiology deals with the study of Viruses. Viruses are
very small ultra-microscopic in nature and they are visible through electron microscope.
Viruses are metabolically inert and are completely dependent on host cell for
replication. Viruses are capable to infect all types of cells from a bacteria to a human.
It contain only one type of nucleic acid that is either DNA or RNA.

Protozoology is a branch of microbiology that deals with the study of protozoans

Immunology is a branch of microbiology that deals with the study of Immune system
of all organism specially human being and animals. In this branch of microbiology the
relationships between host body, pathogen and immunity is studied.

Parasitology is a branch of biology that deals with the study of parasites. This branch
mainly include the study of three major group of bacteria parasitic protozoa, parasitic
worms, and arthropods. In the relationship between host and parasite is also studied.
This parasites may be unicellular or multi-cellular. This parasites are mainly
responsible for causing infection in humans and animals.

(on the basis of Integrative characteristics)

 Microbial cytology
 Microbial physiology
 Microbial genetics
 Microbial ecology
 Microbial taxonomy
 Cellular Microbiology
 Molecular Microbiology

2. Applied Microbiology:

Applied Branches of microbiology are as follows

 Air Microbiology
 Water Microbiology
 Sewage Microbiology

Microbiology Course Pack 10

  Soil Microbiology
 Food microbiology
 Milk Microbiology
 Industry Microbiology
 Medical Microbiology
 Geomicrobiology
 Biotechnology

Air Microbiology
Micro-organism spread through air from one place to another. The micro-organism are
present everywhere in nature and the micro-organism present in air are responsible for
contamination of food or transmission of disease. The diseases such as tuberculosis,
Influenza , and some plant and animal fungal disease etc. spread via air. So it is
important to study specific area for controlling and preventing the spread of some
microbes and this all studies related air and microbes is carried out in this branch.

Water Microbiology
Water is the most important thing that is required for a life form to survive. So the water
used should pure , free of chemicals as well as free from disease causing organisms.
Their are Municipal water purification units that are used to purify water and this quality
of water is checked by examination of water. Water microbiology is used here to check
whether the water is potable or not. Supply of good quality of water is very important
as water can be a source for transmission of various disease.

Sewage Microbiology
Sewage water is the used water of the community. This sewage water contain different
chemicals as well as pathogenic or non-pathogenic micro-organism. If the sewage water
is kept untreated before disposal it can cause harm to the environment as well as any
life form. In sewage microbiology the sewage water is treated by use of some
techniques, chemicals and some useful bacteria.

Soil Microbiology
Soil is supposed to be source of many micro-organism. Study of micro-organism
present is soil is called as soil microbiology. The soil becomes fertile if their are useful
micro-organism present in soil. These micro-organism involve transformation of
elements important for growth of crops as well as degradation of organic matter to
simple form. The bacteria also help in fixation of atmospheric nitrogen for growth of
plants. The soil is a major source of antibiotic producing micro-organism. In soil we
can find variety of micro-organism like bacteria, fungi, viruses and protozoa. Thus
study of soil microbiology is very important.

Food Microbiology
Food microbiology is a important branch as it deals with study of association of micro-
organism with food. Micro-organism deal with food in two ways one it can spoil the
food and spread the infection or disease. Secondly it can used some substrate and
convert into a product that is fermentation. Micro-organism can ferment the substrate
and form the product like curd, Idli , Cheese, Butter etc. If the quality of food is not

Microbiology Course Pack 11

check properly it may result in spread of food born diseases. So the study related
fermentation and quality check of food is important

Milk Microbiology
Milk is a rich and one of the best food and so their are chances of contamination of
milk. If the milk get contaminated their are great chances of spoilage of milk. Milk is
used on large scale in dairy industry where various products like cheese, butter, ghee,
curd are prepared. Useful bacterial present in milk helps in preparation of various milk
product where as harmful bacteria can spoil everything. The food born diseases can
spread through milk if it is not sterilised properly. So it is important to study sterilization
of milk, preparation of various milk product as well as prevention of milk born diseases.

Industrial Microbiology
Industrial microbiology is a branch of microbiology where large amount of substrate
are converted to economically important product by use of micro-organism. Micro-
organism are used in many industries on large scale for production of various product
like antibiotics, beverages, Vaccines, proteins as well as food industries. Therefore the
study of such organism that are be useful in industries for production of economically
important product is very important

Medical Microbiology
In nature their are many useful micro-organism as well as many harmful micro-
organism. Harmful in sense disease causing micro-organism. In this branch the study is
carried out on the causative agent of disease, Identification, pathogenecity, prevention
and cure of disease. It also deals with antibiotics.

Geomicrobiology
This branch of microbiology studies the relation of microbes with some geological
substances like formation of coal, mineral and gas formation as well as recovery of
minerals from low grade ores. It also helps in degradation of hydrocarbon accumulated
in nature in form of pollutants. Micro-organism are used for removal of oil spills in the
ocean by degradation of hydrocarbon and save the aquatic life near the oil spill.

Biotechnology
Biotechnology is a branch of biology that deals with the study of technology and any
life form like microbes, plants animals in the sense to improve quality of life and
develop new technology. It involves techniques like plant tissue culture. Animal tissue
culture, Recombinant DNA technology And genetic modification of organism.

Scope of microbiology

1. Ecology and environment

 Bacteria are primary decomposers – recycle nutrients back into the environment
(sewage treatment plants)
 Winogradsky and M. Beijerinck studied soil microbes and their role in the
biochemical cycles of sulfur, carbon, nitrogen etc.

Microbiology Course Pack 12

  Insect Pest Control: some bacteria are used as bio-pesticides to control Insects
pest. Eg. Bacillus thuringiensis
 Bioremediation: microbes are also used to clean up pollutants and toxic wastes.
Eg. Pseudomonas putida; used to remove petroleum spill.

2. Food microbiology

 Microbes are used in various food and dairy industries to produce various food
products
1. cheese, pickles, sauerkraut, green olives
2. yogurt, soy sauce, vinegar, bread
3. Beer, Wine, Alcohol
 Pasteur (1856) describe fermentation technology

3. Medicine: clinical and pharmaceutical microbiology

 Disease Treatment: Microbes are used to produce Antibiotics

 Eg. Penicillium notatum (Penicillin); discovered by Alexander Fleming (1928)
 Bacteria also synthesize vitamins which is needed for our body.
 Example: E. coli
 Vitamin-B; needed for metabolism
 Vitamin-K; needed for blood clotting
 Gene therapy for treatment of genetic dsiseases

4. Vaccine and immunology

 Vaccine activates immune response.

 Edward Jenner inoculated people with cowpox to protect against smallpox.
 Pasteur developed the rabies vaccine (1885).
 Von Behring and Kitasato (1890) produced toxoid vaccine against diphtheria
and tetanus.
 Metchnikoff (1884) described role of phagocytic cell in defense.

5. Genetic engineering

 Microorganisms are used in Recombinant DNA Technology or Genetic

Engineering to manipulate their gene for the production of useful products such
as enzymes, hormones, interferons, etc
 Microorganisms are used as model organism in molecular biology.

6. Biochemistry and physiology

 Microorganisms are used as a model for study of many biochemical and

physiological processes.

7. Industrial microbiology

Microbiology Course Pack 13

  Microbes are used in economic and industrial purposes.
 Biotechnology, fermentation technology, food and beverages etc are now
established on large industrial scale for income.
 Patients right of procedure

8. Agriculture microbiology

 Genetic engineering is used for the production of transgenic plants and animals.
 Animal and plant improvement by biotechnology for better production, resistant
to environmental fluctuation.
 Molecular farming: transgenic animal or plant are used as bioreactor for mass
production

9. Geochemical microbiology

 Bioleaching: recovery of minerals from low grade ores

 Role of microorganism in geochemical cycle

10. Exomicrobiology

 Exploring for life in outer space

Application

Make a concept map, interconnecting all the topics you learned

from abstraction.

Congratulations! You have finished Lesson 2. Now, you are already prepared
to move to Lesson 3 about Microscopic Methods. Enjoy and keep reading!

Microbiology Course Pack 14

 Lesson 3. Microscopic Methods

Learning Outcomes

At the end of the lesson, you should be able to:

1. Name and identify the parts of the microscope.
2. Relate the function of the different parts of microscope.
3. Demonstrate how to use and care the microscope.
4. Differentiate the types of microscopes.
Time Frame: 1 hour

Introduction

Welcome to Lesson 3
In this lesson, you will learn the parts, function and types of microscopes. In
microbiology subject, microscope is very important equipment to use in order to see
the small microbes.

Activity

What do you know already?

I assume that you learn already the parts of microscope in your previous subjects. Can
you enumerate the different parts of microscope.
Open the link below, watch and listen the video, then answer the question in the analysis
part.
https://www.youtube.com/watch?v=RdfqcWY4E08
https://www.youtube.com/watch?v=pujis3QsA_I

Analysis

What did you learn while watching and listening the video?

Abstraction

Microbiology Course Pack 15

Please read carefully!
A microscope is an instrument that is used to magnify small objects. Some
microscopes can even be used to observe an object at the cellular level, allowing
scientists to see the shape of a cell, its nucleus, mitochondria, and other organelles.
Modern microscope has many parts, the most important pieces are its lenses. It is
through the microscope’s lenses that the image of an object can be magnified and
observed in detail. A simple light microscope manipulates how light enters the eye
using a convex lens, where both sides of the lens are curved outwards. When light
reflects off of an object being viewed under the microscope and passes through the lens,
it bends towards the eye. This makes the object look bigger than it actually is.

Compound microscope consists of at least two lenses, was invented in 1590 by Dutch
spectacle-makers Zacharias and Hans Jansen. Some of the earliest microscopes were
also made by a Dutchman named Antoine Van Leeuwenhoek. Leeuwenhoek’s
microscopes consisted of a small glass ball set inside a metal frame. He became known
for using his microscopes to observe freshwater, single-celled microorganisms that he
called “animalcules.”

Some older microscopes had only one lens, modern microscopes make use of multiple
lenses to enlarge an image. There are two sets of lenses in both the compound
microscope and the dissecting microscope (also called the stereo microscope). Both of
these microscopes have an objective lens, which is closer to the object, and an eyepiece,
which is the lens you look through. The eyepiece lens typically magnifies an object to
appear ten times its actual size, while the magnification of the objective lens can vary.
Compound microscopes can have up to four objective lenses of different
magnifications, and the microscope can be adjusted to choose the magnification that
best suits the viewer’s needs. The total magnification that a certain combination of
lenses provides is determined by multiplying the magnifications of the eyepiece and the
objective lens being used. For example, if both the eyepiece and the objective lens
magnify an object ten times, the object would appear one hundred times larger.

The dissecting microscope provides a lower magnification than the compound

microscope, but produces a three-dimensional image. This makes the dissecting
microscope good for viewing objects that are larger than a few cells but too small to see
in detail with the human eye. The compound microscope is typically used for observing
objects at the cellular level.

Theoretical Principles of Microscopy

To basic principles of microscopy are: magnification, resolution, numerical

aperture, illumination, and focusing.

Magnification

Enlargement or magnification of a specimen is the function of a two-lens

system; the ocular lens is found in the eyepiece, and the objective lens is
situated in a revolving nose-piece. These lenses are separated by the body

Microbiology Course Pack 16

tube. The objective lens is nearer the specimen and magnifies it, producing
the real image that is projected up into the focal plane and then magnified by
the ocular lens to produce the final image.

The most commonly used microscopes are equipped with a revolving

nosepiece containing four objective lenses possessing different degrees of
magnification. When these are combined with the magnification of the ocular
lens, the total or overall linear magnification of the specimen is obtained.

Resolving Power or Resolution

Although magnification is important, you must be aware that unlimited

enlargement is not possible by merely increasing the magnifying power of the
lenses or by using additional lenses, because lenses are limited by a property
called resolving power. Resolving power is the ability of a lens to show two
adjacent objects as discrete entities. When a lens cannot discriminate, that is,
when the two objects appear as one, it has lost resolution. Increased
magnification will not rectify the loss, and will, in fact, blur the object. The
resolving power of a lens is dependent on the wave-length of light used and
the numerical aperture, which is a characteristic of each lens and imprinted
on each objective. The numerical aperture is defined as a function of the
diameter of the objective lens in relation to its focal length. It is doubled by
use of the substage condenser; which illuminates the object with rays of light
that pass through the specimen obliquely as well as directly. Thus, resolving
power is expressed mathematically, as follows:

Resolving power = Wavelength of Light .

2 (Numerical Aperture)

Based on this formula, the shorter the wave-length, the greater the resolving
power of the lens. Thus, short wavelengths of the electromagnetic spectrum
are better suited than longer wavelengths in terms of the numerical aperture.

However; as with magnification, resolving power also has limits. You might
rationalize that merely decreasing the wavelength will automatically increase
the resolving power of a lens. Such is not the case, because the visible portion
of the electromagnetic spectrum is very narrow and borders on the very short
wavelengths found in the ultraviolet portion of the spectrum.

The relationship between wavelength and numerical aperture is valid only for
increased resolving power when light rays are parallel. Therefore, the
resolving power is dependent on another factor, the refractive index. This is
the bending power of light passing through air from the glass slide to the
objective lens. The refractive index of air is lower than that of glass, and as

Microbiology Course Pack 17

light rays pass from the glass slide into the air, they are bent or refracted so
that they do not pass into the objective lens. This would cause a loss of light,
which would reduce the numerical aperture and diminish the resolving power
of the objective lens. Loss of refracted light can be compensated for by
interposing mineral oil, which has the same refractive index as glass, between
the slide and the objective lens. In this way, decreased light refraction occurs
and more light rays enter directly into the objective lens, producing a vivid
image with high resolution.

Illumination

Effective illumination is required for efficient magnification and resolving

power. Since the intensity of daylight is an uncontrolled variable, artificial
light from a tungsten lamp is the most commonly used light source in
microscopy. The light is passed through the con-denser located beneath the
stage. The condenser contains two lenses that are necessary to produce a
maximum numerical aperture. The height of the condenser can be adjusted
with the con-denser knob. Always keep the condenser close to the stage,
especially when using the oil-immersion objective.

Between the light source and the condenser is the iris diaphragm, which can
be opened and closed by means of a lever; thereby regulating the amount of
light entering the condenser. Excessive illumination may actually obscure the
specimen because of lack of contrast. The amount of light entering the

Microbiology Course Pack 18

microscope differs with each objective lens used. A rule of thumb is that as
the magnification of the lens increases, the distance between the objective lens
and slide, called working distance, decreases, whereas the numerical aperture
of the objective lens increases.

How to Use and Care the Microscopes (open and study the link below)

https://www.austincc.edu/microbugz/handouts/Microscope%20Use%20and%20Care.
pdf

Microscope Care
Proper care and maintenance of your microscope can extend its life by many years.

Handle with care

Most microscope problems occur as a result of improper handling. When carrying your
microscope, hold it by the base and the metal support arm. Do not pick it up by the
stage, as this can cause misalignment. When transporting it, use a microscope bag.

Keep lenses clear of slides

When using your microscope and adjusting the focus you will need to lower the
objective lens down as far as it will go. However, you should never allow the lens to
touch the slide you are looking at. Dirty lenses can be difficult to clean.

Clean after using immersion oil

If using immersion oil, always ensure the objectives are cleaned immediately after use.
Objective, eyepieces and condenser may be removed for cleaning. Use only lens paper
and lens cleaner. Do not use solvents.

Cover when not in use

All microscopes are sold with dust covers. Always keep your microscope covered when
not in use even if the microscope is stored in a cabinet. Eye tubes also need to be kept
free of dust so do not store a microscope without the eyepieces. If the microscope
eyepieces must be removed, cover the tubes with caps or a plastic bag with a rubber
band around the eye tube.

Look after the bulb

After using the microscope, turn off the illuminator and wait for it to cool for several
minutes before putting it away. By allowing the bulb to cool you will extend its life.

Microbiology Course Pack 19

When turning the microscope on and off, use the switch not the power point. Do not
switch the microscope on while using full light intensity. Never touch the bulb with
your fingers as the body oils can burn into the bulb and reduce its life. Use a tissue.
Keep a store of replacement bulbs and always use the correct bulb.

Store in a clean, dry place

Make sure you do not store your microscope in an area that has corrosive chemical
fumes that can destroy lenses or metal parts or beside solutions that may leak. Salt air
and pervasive damp can also cause damage over time. Make sure your cabinet is
ventilated.
Only use special lens paper or wipes for cleaning the lenses
Microscope lenses can easily be scratched and should be treated with great care. Use
an aspirator to remove dust. Sticky residue can be removed with lens paper moistened
with distilled water or lens cleaning solution and rubbed gently using a circular motion.
Never use sharp instruments or anything abrasive on the microscope lenses.

Keep your User's Manual and wrenches in a safe place

Each microscope should come with a user's manual and specialist wrenches as required.
Always consult the User's Manual before making any adjustments to your microscope
and use the wrenches provided. Never over-tighten or use force when performing any
maintenance on your microscope, or use inappropriate tools. This can damage the parts.

Perform an annual maintenance check

On an annual basis moving parts on the microscope should be cleaned and lubricated.
Clean grease and dirt from sliding surfaces using a clean cloth. Apply a very thin layer
of lithium-based grease to the sliding surfaces. Do not grease the teeth of the rack and
pinion gears. Inspect the power cords and plugs for safety and stock up on a supply of
replacement bulbs.

Have your microscope serviced professionally

A rule of thumb for frequency of servicing is every 200 hours of use or every 3 years,
whichever comes first.

Types of Microscopes
1. Stereo Microscope
2. Compound Microscope
3. Inverted Microscope
4. Metallurgical Microscope
5. Polarizing Microscope

Microbiology Course Pack 20

 Stereo Microscope

Stereo microscope is used to look at a variety of samples that you would be able to
hold in your hand. A stereo microscope provides a 3D image or "stereo" image and
typically will provide magnification between 10x - 40x. The stereo microscope is used
in manufacturing, quality control, coin collecting, science, for high school dissection
projects, and botany. A stereo microscope typically provides both transmitted and
reflected illumination and can be used to view a sample that will not allow light to pass
through it.
Compound Microscope

A compound microscope may also be referred to as a biological microscope.

Compound microscopes are used in laboratories, schools, wastewater treatment plants,
veterinary offices, and for histology and pathology. The samples viewed under a
compound microscope must be prepared on a microscope slide using a cover slip to
flatten the sample. Students will often view prepared slides under the microscope to
save time by eliminating the slide preparation process.

The compound microscope can be used to view a variety of samples, some of which
include: blood cells, cheek cells, parasites, bacteria, algae, tissue, and thin sections of
organs. Compound microscopes are used to view samples that cannot be seen with the
naked eye. The magnification of a compound microscope is most commonly 40x, 100x,
400x, and sometimes 1000x. Microscopes that advertise magnification above 1000x
should not be purchased as they are offering empty magnification with low resolution.

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 Inverted Microscope

Inverted microscopes are available as biological inverted microscopes or metallurgical

inverted microscopes. Biological inverted microscopes provide magnification of 40x,
100x and sometimes 200x and 400x. These biological inverted microscopes are used to
view living samples that are in a petri dish. An inverted microscope allows the user to
place the petri dish on a flat stage, with the objective lenses housed beneath the stage.
Inverted microscopes are used for in-vitro fertilization, live cell imaging,
developmental biology, cell biology, neuroscience, and microbiology. Inverted
microscopes are often used in research to analyze and study tissues and cells, and in
particular living cells.

Metallurgical Microscope

Metallurgical inverted microscopes are used to examine large parts at high

magnification for fractures or faults. They are similar to biological inverted microscope
in the magnification provided, but one primary difference is that the samples are not
placed in a petri dish, but rather a smooth side of the sample must be prepared so it can
lay flat on the stage. This smooth sample is polished and is sometimes referred to as a
puck.

Metallurgical microscopes are high power microscopes designed to view samples that
do not allow light to pass through them. Reflected light shines down through the
objective lenses providing magnification of 50x, 100x, 200x, and sometimes 500x.
Metallurgical microscopes are utilized to examine micron level cracks in metals, very
thin layers of coatings such as paint, and grain sizing.

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Metallurgical microscopes are utilized in the aerospace industry, the automobile
manufacturing industry, and by companies analyzing metallic structures, composites,
glass, wood, ceramics, polymers, and liquid crystals.

Polarizing Microscope

Polarizing microscopes use polarized light along with transmitted and, or reflected
illumination to examine chemicals, rocks, and minerals. Polarizing microscopes are
utilized by geologists, petrologists, chemists, and the pharmaceutical industry on a daily
basis.

All polarizing microscopes have both a polarizer and an analyzer. The polarizer will
only allow certain light waves to pass through it. The analyzer determines the amount
of light and direction of light that will illuminate the sample. The polarizer basically
focuses different wavelengths of light onto a single plane. This function makes the
microscope perfect for viewing birefringent materials.

Essential Features of other Microscopes

Brightfield Microscope

This instrument contains two lens systems for magnifying specimens: the ocular lens in
the eyepiece and the objective lens located in the nosepiece. The specimen is
illuminated by a beam of tungsten light focused on it by a sub-stage lens called a
condenser, and the result is that the specimen appears dark against a bright background.
A major limitation of this system is the absence of contrast between the specimen and
the surrounding medium, which makes it difficult to observe living cells. Therefore,
most brightfield observations are performed on nonviable, stained preparations.

Darkfield Microscope

This is similar to the ordinary light microscope; however, the condenser system is
modified so that the specimen is not illuminated directly. The con-denser directs the
light obliquely so that the light is deflected or scattered from the spec-imen, which then

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appears bright against a dark background. Living specimens may be observed more
readily with darkfield than with brightfield microscopy.

Phase-Contrast Microscope

Observation of microorganisms in an unstained state is possible with this microscope.

Its optics include special objectives and a condenser that make visible cellular
components that differ only slightly in their refractive indexes. As light is transmitted
through a specimen with a refractive index different from that of the surrounding
medium, a portion of the light is refracted (bent) due to slight variations in density and
thickness of the cellular components. The special optics convert the difference between
transmitted light and refracted rays, resulting in a significant variation in the intensity
of light and thereby producing a discernible image of the structure under study. The
image appears dark against a light background.

Fluorescent Microscope

This microscope is used most frequently to visualize specimen that are chemically
tagged with a fluorescent dye. The source of illumination is an ultraviolet (UV) light
obtained from a high-pressure mercury lamp or hydrogen quartz lamp. The ocular lens
is fitted with a filter that permits the longer ultraviolet wavelengths to pass, while the
shorter wavelengths are blocked or eliminated. Ultraviolet radiations are absorbed by
the fluorescent label and the energy is re-emitted in the form of a different wavelength
in the visible light range. The fluorescent dyes absorb at wavelengths between 230 and
350 nanometers (nm) and emit orange, yellow, or greenish light. This microscope is
used primarily for the detection of antigen-antibody reactions. Antibodies are
conjugated with a fluorescent dye that becomes excited in the presence of ultraviolet
light, and the fluorescent portion of the dye becomes visible against a black background.

Electron Microscope

This instrument provides a revolutionary method of microscopy, with magnifications

up to one million. This permits visualization of sub-microscopic cellular particles as
well as viral agents. In the electron microscope, the specimen is illuminated by a beam
of electrons rather than light, and the focusing is carried out by electromagnets instead
of a set of optics. These components are sealed in a tube in which a complete vacuum
is established. Transmission electron microscopes require specimen that are thinly
prepared, fixed, and dehydrated for the electron beam to pass freely through them. As
the electrons pass through the specimen, images are formed by directing the electrons
onto photographic film, thus making internal cellular structures visible. Scanning
electron microscopes are used for visualizing surface characteristics rather than
intracellular structures A narrow beam of electrons scans back and forth, producing a
three-dimensional image as the electrons are reflected off the specimen's surface.

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 Application

https://www.pobschools.org/cms/lib/NY01001456/Centricity/
Domain/349/TheMicroscope-howtouse.pdf
Open this link and answer page 3

Congratulations! You did a great Job! You have finished this

module.

Assessment will be part of the summative test

Module Summary
It is hoped that after finishing this module, you are now more familiar about the
different concepts and principles of Microbiologyt. It is assumed also that you know
already the scope and branches of Microbiology. It is hoped also that you familiarized
the different parts and types of microscopes. Congratulations!
To sum up Module 1 Highlights:
 Microbiology is a vast and multidisciplinary field which overlaps with other life
sciences such as genetics, biochemistry, molecular biology and even engineering.
The science of microbiology aims to gain and expand our fundamental
understanding of microorganisms by studying their morphology, metabolism,
physiology, reproduction and genetics, while others investigate their interactions
with other organisms and role in ecology.

 Generally microbes can be divided into two categories: the cellular microbes (or
organisms) and the acellular microbes (or agents). In the cellular camp we have the
bacteria, the archaea, the fungi, and the protists (composed of algae, protozoa, slime
molds, and water molds). Cellular microbes can be either unicellular, where one
cell is the entire organism, or multicellular, where hundreds, thousands or even
billions of cells can make up the entire organism. In the acellular camp we have the
viruses and other infectious agents, such as prions and viroids.

 The scope in this field is very important due to the involvement of microbiology in many
fields like medicine, pharmacy, diary, industry, clinical research, water industry, agriculture,
chemical technology and nanotechnology. The study of microbiology contributes greatly to
the understanding of life through enhancements and intervention of microorganisms.

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REFERENCES
Colbert, Bruce. J. . 2016. Microbiology: Practical Applications and Infection Prevention.
Boston, MA. Cengage Learning.

Cowan, M. .2015. Microbiology. A System Approach.4th Ed. New York. MacGraw-Hill

Education.

Engelkiek, Paul G. 2015. Burtons’s Microbiology for Health Sciences. 10th Ed.
Philadelphia: Wolters Kluwer.

Lewis, J. D..2013. Marine Microbial Ecology. New Delhi. Random Exports.

Lunardi, M. Alfieri, A..2016. Microbiology and Virology. New York, NY: Magnum
Publishing

Tortora, Gerald J. Funke, Berdell, R. & Case, Christine, L. .2011. Microbiology: An

Introduction. 10th Edition. Pearson Education, Inc. Benjamin Cummings.

Web Links
https://www.sciencedirect.com/book/
https://.britanica,com/science/biology#ref498653
https://www.bookdepository.com/campbell-biology-lisa-urry/
https://www.sciencefacts.net/wp-content/uploads/2020/06/Cell-Membrane-Structure-
Diagram-jpg
http://serendip.brynmawr.edu/sci_edu/waldron/pdf/BreathingLabProtocol.pdf

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