The awareness of
A HAND AT MICROSCOPY
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contamination due to
microorganisms in the
Thando Sgalelana
environmentPractical 1
[Course title]
Practical 1
II
Table of Contents
TABLE OF CONTENRS: ……………………………………………………………………………………...II
LIST OF FIGURES: …………………………………………………………………………………………...III
ABSTRACT: ......................................................................................................................................................... 2
List Of Figures: ..................................................................................................................................................... 1
ABSTRACT: ......................................................................................................................................................... 2
AIM ........................................................................................................................................................................ 7
Objectives .............................................................................................................................................................. 7
Materials: .............................................................................................................................................................. 8
Methods: ................................................................................................................................................................ 7
Results: .................................................................................................................................................................. 8
Discussion: ................................................................................................................Error! Bookmark not defined.
References/bibliography:………………………………………………………………………………………..8
List Of Figures:
FIGURE 2: BACTERIA AND ARCHAEA....................................................................................................................... 2
FIGURE 3: SHAPES OF BACTERIA............................................................................................................................. 3
FIGURE 4: FUNGI..................................................................................................................................................... 3
FIGURE 7: RHIZOPUS STOLONIFER .......................................................................................................................... 3
FIGURE 5: YEAST FINGI ........................................................................................................................................... 3
FIGURE 6:MUSHROOM FUNGI................................................................................................................................ 3
FIGURE 6: MUSHROOM FUNGI ............................................................................................................................... 3
FIGURE 8: PROTOZOA ............................................................................................................................................. 3
FIGURE 9: SLIME MOLD .......................................................................................................................................... 4
FIGURE 10: PROTISTS CATEGORIES ......................................................................................................................... 4
FIGURE 11: TOMATOES INFECTED BY A VIRUS ........................................................................................................ 4
FIGURE 12: VIRUS ................................................................................................................................................... 4
TABLE 1: A TABLE SHOWING THE DIFFERENT TYPES OF COLONIES AND THE NUMBER OF COLONIES FORMED ................ 8
ABSTRACT; EXPERIMENT 1:
Microorganisms are small organisms, too small to be seen with the naked eye, they lack
highly differentiated cells and distinct tissues. Microorganisms are omnipresent. Microbes
include Bacteria, Fungi, and Viruses. However, scientists have produced microbiological
techniques to grow microbes through a process of Inoculation. Inoculation is accomplished
with the use of an agar medium surface. Bacterium can multiply and form a colony that can
be seen with the naked eye. Students collected samples in the laboratory and deposited the
samples to the growth media and performed a follow up, observing the colonies that had
formed on the agar plates. The aim was to prove that microorganisms exist everywhere in our
environment and how easily substances like media, pure specimens, food, and water can be
contaminated and why it is important to work aseptically in the laboratories to avoid
contamination.
INTRODUCTION; EXPERIMENT 1
Microbes form a large part of the biosphere because they are omnipresent and can be
beneficial or unfavourable to humans. Microbes infiltrate two groups, prokaryotes, and
eukaryotes.
Prokaryotic cells lack the membrane-bound organelles this group includes viruses, bacteria
and archaea and are unicellular. Eukaryotic cells have a nucleus and many other membranebound organelles that separate some cellular material and process from others. This group
includes algae, fungi, protists and animal cells that have a nucleus.
Figure 1: Bacteria {www.visiblebody.com}
Bacteria and Archaea
Figure 2: Bacteria and Archaea
Bacteria and Archaea are small unicellular organisms.
Their cell size ranges from 0.5 to 1.0 micrometre in
diameter. Bacteria and Archaea have many cell shapes and
contain a circular DNA. These microbes reproduce
asexually (binary fission is one of the reproduction
methods). Microbes that inhabit the large intestine help
keep the body digest food and produce vitamins. Archaea
{slideplayer.com}
2
are more related to eukaryotes than the bacteria. (Willey, 2013)
Figure 3: Shapes of bacteria
{gilbertlab.com}
Fungi
Figure 4: Fungi
{careerpower.in}
Fungi are Eukaryotes. Fungi include moulds, yeasts, and mushrooms. The hyphae are the
main parts of fungi and consist of mycelium. Fungi can reproduce sexually and asexually.
Some of the beneficial roles are bread raising(yeast), production of antibiotics, and
decomposition of dead organisms.
Protista
Figure 8: Protozoa
{researchgate.net}
Figure 5: Rhizopus stolonifer
{sciencekids.in}
Figure 6: Yeast fingi
Figure
Figure 6:Mushroom
7: Mushroomfungi
fungi
{globalgarden.co}
3
{sciencephoto.com}
These are unicellular eukaryotes, but they lack a cell wall. Protist were the first Eukaryotes to
evolve. Some protists are photosynthetic. These include algae, amoebas, and ciliates.
Primarily aquatic organisms that absorb free nutrients from the environment, some live as
parasites whilst others as free entities.
Figure 10: Protists categories
Figure 9: Slime mold
{pl.pinteresr.com}
{researchgate.net}
Virus
Figure 12: Virus
Figure 11: Tomatoes infected by a virus
{researchgate.net}
{synngenta.co.in}
Viruses are the smallest pathogens that require a host cell to survive and reproduce. Some
viruses are enveloped, others are non-enveloped. Plant viruses affect important crops such as
tobacco and tomatoes. Animal viruses cause deadly diseases. (Singh, 2016)
4
ABSTRACT; EXPERIMENT 2:
Microscopy is a technical field in which a microscope is used to view samples and
specimens that cannot be seen with the naked eye, for example microorganisms. A
Microscope is an Optical Instrument that is used for microscopic(very small objects), such as
animal or plant cells, or mineral samples. There are different types of microscopes, here are 5
types:
1. Compound Microscope
It is an instrument that has two lenses (set of two lenses) these lenses is objectives and
ocular. Furthermore, they use visible light as a source of illumination.
2. Darkfield Microscope
These microscopes have a device that scatters light from the illuminator. In addition,
it does this to make the specimen appear white against the black background.
3. Electron Microscope
It is a scope that instead of light uses a flow of electron to produce an image.
Moreover, this microscope enhances the images of viruses, protein, lipids, ribosomes,
and even small molecules.
4. Fluorescence Microscope
These scopes use ultraviolet light to illuminate specimens that fluoresce. Besides,
mostly, a fluorescent antibody or dye is added on the viewed specimen.
5. Contrast/Phase Microscope
This scope uses a special condenser that allows the examination of structures inside
the cells. Also, they use a compound light. Furthermore, these microscopes take
advantage of different refractive indexes for the examination of live organisms.
In addition, the final image produced by these microscopes is a combination of light
and dark.
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INTRODUCTION; EXPIREMENT 2
Microscopes are in different fields for different purposes, here are 5 that are most commonly
used:
Tissue Analysis
This study of tissues and organs through sectioning, staining, and examining the sections
under a microscope is called Medical Histology, also known as, microscopic anatomy and
Histochemistry. Histology allows for the visualization of the tissue structure and the
characteristic changes that the tissue may have undergone.
{National Library of Medicine, national center for biotechnology information. STAT
PEARLS textbook – History, Staining by Tatyana S. Gurina; Lary Simms}
[ https://www.ncbi.nlm.nih.gov/books/NBK557663/ ]
Examination of Forensic Evidence
Electron microscopy (EM) is sued in forensic investigations. Many micro-traces found at
crime scenes such as, glass and paint fragments, tool marks, drugs, explosives, and Gunshot
Residue (GSR), to name a few, can be visually and chemically analyzed with Scanning
Electron Microscopy (SEM). Forensic Microscopist uses microscopes to locate, recover,
identify, and compare trace evidence.
{Banaras Hindu University - Microscopes in Forensic Science introduction. pdf}
[https://www.bhu.ac.in/Content/Syllabus/Syllabus_300620200523100800.pdf]
The Study of the Role of Protein Within the Cell
An Electron Microscope is used to visualize protein molecules. It generates high-energy
electrons which in turn gives an electronic image to observe protein molecules. A technique
called Fluorescent Analog Cytochemistry is used to visualize any protein inside a cell, in
which a purified protein is coupled to a fluorescent dye and micro-injected into a cell. In this
way, the journey of the injected protein can be followed in a fluorescence microscope as the
cell grows and divides.
{National Library of Medicine, national center for biotechnology information. Molecular
Biology of the Cell. 4th edition – Visualizing Molecules in Living Cells. By Alberts B,
Johnson A, Lewis J}
[https://www.ncbi.nlm.nih.gov/books/NBK26893/]
{toppr from BYJU’S, Microscope-Definition, Type, Uses, Parts}
[https://www.toppr.com/guides/biology/microbiology/microscope-types-uses-parts/]
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AIM & OBJECTIVE; EXPIREMENT 1
AIM
The aim of this experiment is to prove that microorganisms are EVERYWHERE in the
environment, whether it be in the air, the surfaces, and our bodies.
Objectives
1. To investigate the presences of microorganisms in the environment.
2. To show how easily media surface and other substances can be contaminated.
3. To call attention to the necessity of working aseptically in the laboratory to avoid such
contamination.
Methods; Experiment 1:
Lab coats were worn immediately upon entering the laboratory. Once at the lab station, 4
Agar Plates Prepared were labelled with the samples that were collected from the air, bench,
and washed and unwashed hands. One plate labelled “air” was exposed to the laboratory
atmosphere for 30 minutes. A cotton swab was used to wipe the lab bench and flecked on the
entire surface of the second plate labelled “bench”, (cotton swab container was broken, this
was an indication that it has been used). The surface of the third plate labelled “unwashed
hands” was pushed with fingers. The last plate was touched with fingers after washing hands
with a fair amount of antibacterial soap and water and dried dry. All the plates were then
incubated for 24 hours at 37 degrees Celsius. A follow up practical was done after 24 hours
and the results we recorded.
AIM & OBJECTIVE; EXPIREMENT 2
AIM
The aim of this experiment was to not just familiarize ones-self to the microscope, but to also
be able to view and identify the type of bacteria being observed under the microscope.
Objectives
1. To familiarize ourselves with the functions of a microscope.
2. To identify types of bacteria, through the shapes of their cells.
3.
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Materials and Method; Experiment 2:
1.
2.
3.
4.
A Binocular Compound Light Microscope.
Prepared and stained slides with microbial specimens.
Lens cleaning tissue paper.
Immersion Oil.
Materials:
1.
2.
3.
4.
5.
4X Petri Dishes filled with a type of Culture Media called Ager
1X Cotton Swab
A bottle of antibacterial soap
Water
Paper towel
Methods:
Results:
A colony is visible microbial growth on a solid nutrient medium and is made up of the same
type of microbes. It reproduces through binary fission.
Plates were taken out the incubator and using a marker, the total number of colonies were
counted, and the types of colonies were recorded.
Table 1: A table showing the different types of colonies and the number of colonies formed
Bench
Air
Washed
Unwashed
Colonies
10 colonies
10 colonies
128 colonies
Shape
Texture
Colour
Elevation
Punctiform
Dry
1 yellow, 9 buff
Flat
7 colonies + 1
large cluster of
colonies
Irregular
Rough
1 yellow, 7 buff
Raised
Punctiform
Dry
Buff
Flat
Circular
Smooth
Buff
Convex
Discussion:
In this experiment, the aim was to show how easily media and other substances such as food,
water, and pure specimens can contaminated and why it is necessary to work aseptically in
the laboratory to avoid such contamination. The experiment proved that microorganisms are
8
literally everywhere, as seen in the results. The hypothesis stated that microorganisms exist
everywhere. This was strongly matched as all the agar plates had growing colonies on their
surface.
This experiment generated evidence within 24 hours after it was performed. Because of this,
the results of the experiment were accurate to the hypothesis stated. The results were different
for each different specimen. Comparing between washed and unwashed hands agar plate
results, there were numerical colonies present on the unwashed agar media than that of the
washed hands agar media. It was clearly seen that we tend to contaminant different kinds of
microorganisms from our environment. After washing our hands with a 99.9% antibacterial
soap, fewer microorganisms were present on our hands. The unwashed hands agar plate has
formed an elevated large cluster of colonies, this showed how we carry a large number of
microorganisms on our hands. That highlighted the importance of washing of hands regularly
because you might not know what kind of microbe you can carrying on your hands, it might
be pathogenic and lead to critical sickness.
Comparing the air and the bench results, air had a greater number of colonies on its agar plate
than that of the bench. This is because the laboratory bench was disinfected with a 75%
ethanol disinfector, providing that it is necessary to disinfect the environment that we are in
close contact with. The air in the laboratory was made up of different gases, some known and
some unknown. The air carried microbes in it that we were not aware of so that is why the
growing colonies were quite numerous on the agar media since the air was not as clean as it
needed to be. The air labelled agar plate had irregular shapes formed in it, this was also an
indication that there are different kinds of microorganisms present in the air.
The time spam that was used (24 hours) was fair because it gave the microbes time to be
visible on the agar media.
The hypothesis was that microorganisms are present everywhere and can be contaminated.
The results accurately support the hypothesis. Microorganisms exist everywhere hence the
importance to practice good hygiene and keep our environment clean and disinfected at all
times and it is necessary to work aseptically in the laboratory to avoid such contamination.
This experiment brought awareness to our account, and it is advisable to educated others with
the knowledge that was acquired during the performance of this experiment.
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10
Results; Experiment 2:
Specimen with blue staining Specimen with red staining
Image captured at 4x
magnification.
Image captured at 10x
magnification.
Image captured at 40x
magnification
Image captured at 100x
magnification
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CONCLUSION:
With the experiment performed, and the results collected, we believe we have reached our
aim in proving that microorganisms are everywhere in the environment at difference
concentrations/intensity.
Our hypothesis is that microorganisms are present everywhere and can contaminate pure
substances and the results we gathered accurately support said hypothesis. Therefore, in
conclusion we would like to bring to your attention the importance to practicing good
hygiene and keep our environment as clean as we possibly can, with products such as
disinfectants, at all times and why it is necessary to work aseptically in laboratories and
healthcare facilities to avoid such contamination. We are pleased to say that this experiment
and report serves as a learning material to educate others with the knowledge that was
acquired during the performance of this experiment.
CONCLUSION; Experiment 2
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References / bibliography
•
Brownstein J. & Chitale R. 2008. 10 Germy surfaces you touch each day. ABC News
Medical Unit
[https://abcnews.go.com/Health/ColdandFluNews/story?id=5727571&page=1]
•
Cambridge Dictionary, 4th ed. 2021. CEF Level: A1-B2. Cambridge: Mclntosh, Colin.
•
https://encryptedtbn0.gstatic.com/images?q=tbn:ANd9GcR9zRQPv8RHBj74WkgrcFPO3sDKmgqjES
q-jg&usqp=CAU
•
https://www.google.com/url?sa=i&url=https%3A%2F%2Fwww.sciencephoto.com%2
Fmedia%2F922083%2Fview%2Fbacteria-in-human-large-intestineillustration&psig=AOvVaw28Fqcl0WdXU90duB2_z4n&ust=1709387030646000&source=images&cd=vfe&opi=89978
449&ved=0CBMQjRxqFwoTCKjEqYiZ04QDFQAAAAAdAAAAABAJ
•
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Figure 4: Fungi; https://images.app.goo.gl/urC5iQ5Sy4nYKvTN9
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Figure 7: Rhizopus stolonifer; https://images.app.goo.gl/BTC3CedPETd2uyFL6
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Figure 6: Mashrooms Fungi; https://images.app.goo.gl/xWoic9NHekTrtAoo9
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Figure 5: https://images.app.goo.gl/ucx7KPB61KQRjvE38
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9. https://images.app.goo.gl/wvLw6cJ2kg9uZkXm8
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10. https://images.app.goo.gl/QHFzh82Nn6ZRAWgp9
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11. https://images.app.goo.gl/FGAuv7BcbhHpUUx68
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12. https://images.app.goo.gl/n4whpR8cW4Tcazra9
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