Laboratory Report in Microbiology and Parasitology

Republic of the Philippines
BATANGAS STATE UNIVERSITY

The National Engineering University
Pablo Borbon
Rizal Avenue Ext., Batangas City, Batangas, Philippines 4200
Tel Nos.: (+63 43) 980-0385; loc 1127
E-mail Address: conahs.ph@g.batstate-u.edu.ph | Website Address: https://www.batstate-u.edu.ph
College of Nursing and Allied Health Sciences
Laboratory Activity No 1:
Manipulating a Microscope
1
Pablo Borbon
Tel Nos.: (+63 43) 980-0385; loc 1127
A. Introduction
Any medical student needs to be how to manipulate a microscope, yet doing so might be
scary. A compound light microscope is the most typical form of microscope you'll use in your
biology labs. This particular type of microscope views tiny things using both visible light and
electrons. The visible light allows you to see what is being magnified while the electrons are
employed to form an image that appears on a screen. You must comprehend how this type of
microscope operates and what the names of its components are in order to use one. Also, you
must be familiar with how to change its settings so that you can observe objects at various
magnifications and how to focus the image so that it is sufficiently clear for identification. A
revolutionary atomic force microscope designed to measure the force required to move samples
adhering to surfaces in the micron range is called a manipulation force microscope. It has
effectively described how proteins and live cells adhere to substrates.
B. Materials
Following materials is used in virtual experiments
• Compound microscope
• Bacterial slides
• Immersion oil
• Lenses paper
• Mobile phone
C. Procedure
1. For a virtual microscope simulation, click the linked link.
2. The website will flash an overview; click "Launch Activity" to begin.
3. You will then be taken to the primary virtual microscope where you can choose an
option the menu below to explore, learn more about the microscope, consult the resource
guide, or test your knowledge.
4. The main activity can be started by clicking the "Explore" bar. A view of the many
laboratories on the screen, tools including the microscope, immersion oil, lens paper, and
the slide catalog will be visible.
5. Get to the microscope's slide catalog by clicking here. A slide catalog including a range
of prepared slides, including sample slides, plant slides, animal slides, bacterial slides,
and the human for you to view, will show up after clicking. But in this lab experiment,
we'll look at the bacterial slides. So, select the next-to-last panel.
6. The slide catalog will reappear after selecting the bacterial slide and contain additional
stained slides, including gram stain mix, endospore stain, and acid-fast mix. Examine
each slide of bacteria.
2
Pablo Borbon
Tel Nos.: (+63 43) 980-0385; loc 1127
7. A microscopic view of the specimen will be displayed on the screen after selecting one
of the prepared slides.
8. Adjust the coarse focus, fine focus, and light after looking at the slide at the lowest
magnification (4x), to get a clear, detailed image of the bacterial sample you are looking
at.
9. Instead of skipping a step, change the magnification in sequence, going higher than the
first magnification to avoid harming the slides and objective lenses.
10. As soon as you reach the 100x magnification, which is the maximum, make sure you
click it to add immersion oil. This is done to improve the microscope's resolution and
create images of excellent quality.
11. After using the 100x magnification, wipe the lens and the slide with the lens paper.
12. Do the techniques once more to the other bacterial slides that are available in the slide
catalog, and then properly inspect them.
D. Data and Analysis
BACTERIAL SLIDES
MAGNIFICATION Acid Fast Endospore Stain Gram Stain Mix
Mix
4x
10x
3
Pablo Borbon
Tel Nos.: (+63 43) 980-0385; loc 1127
40x
100x
Actual Sample from

the Internet
4
Pablo Borbon
Tel Nos.: (+63 43) 980-0385; loc 1127
Description/ Use of • Acid-fast organisms • Endospores • A Gram stain is

The Sample are characterized by staining is the type colored purple. When
wax-like, nearly of staining to the stain combines
impermeable cell recognize the with bacteria in a
walls; they contain presence spore in sample, the bacteria
mycolic acid and bacterial vegetative will either stay purple
large amounts of cells. The bacterial or turn pink or red. If
fatty acids, waxes, endospores need a the bacteria stay
and complex lipids. staining which can purple, they are
Acid-fast organisms penetrate wall Gram-positive. If the
are highly resistant thickness of spore bacteria turn pink or
to disinfectants and bacteria. A method red, they are
dry conditions. of endospores Gramnegative.
• Mycobacterium staining is • A Gram stain is
tuberculosis is Schaeffer Fulton colored purple. When
primarily a lung method that used the stain combines
pathogen; therefore, Malachite Green. with bacteria in a
for all types of • Endospore Staining sample, the bacteria
tuberculosis, the is a technique used will either stay purple
primary specimen in bacteriology to or turn pink or red. If
required in acid-fast identify the the bacteria stay
smear microscopy is presence of purple, they are
sputum endospores in a Gram-positive. If the
bacterial sample, bacteria turn pink or
which can be useful red, they are
for classifying Gramnegative.
bacteria.
5
Pablo Borbon
Tel Nos.: (+63 43) 980-0385; loc 1127
E. Guide Questions
1. What is magnification? What is its importance in microscopy?
A microscope's ability to produce a picture of an object at a scale that is larger (or even
smaller) than its real size is known as magnification. Only when it is feasible to discern more details of an
object in the image than when observing the object with the unaided eye can magnification serve a
beneficial purpose. We wouldn't know anything about minute things like germs and viruses or distant
things like stars and galaxies if we couldn't magnify little items. We use microscopes to examine items
more closely and see details that aren't visible to the naked eye. We wouldn't know about the existence of
cells, how plants breathe, or how rocks evolve over time without them.
2. Based on your activity, list some parts of the microscope that is mostly used in the
experiment? Try to identify its role in the activity. Give at least one example. (Aside from the
microscope you may also refer to other tools and equipment used in the activity.
A microscope slide is a small piece of glass used to hold objects for microscopic
examination. An object is held on a microscope slide, which is a flat, thin piece of glass that is typically
75 by 26 mm (3 by 1 inches) and about 1 mm thick. Usually, the slide and the object are placed into the
microscope together before being mounted (attached) on the slide.
3. When switching lenses in a microscope, why is it important to move through lenses in order?
The ability to change lenses in a microscope is crucial because it enables you to view
objects using lenses with various focal lengths. You can see several things at once as a result.
In order to change lenses, you may either move the objective lens closer to or farther from
your specimen or move the eyepiece lens closer to or farther from your eye. While the latter
enables you to see more clearly, the former allows you to see more detail.
4. For you, what is the most important laboratory safety rule? Justify your answer.
The most essential lab safety rule is to be aware of where safety gear, like a fire
extinguisher, is located and how to use it. Despite all safety measures, there is always a
potential of an accident in a lab. This is due to the fact that human mistake is always possible.
5. If you are to create and invent one lab tool and equipment, what would it be? How will it be
different from the tools available in the laboratory? (You can be creative on this part)
A hand robot that can detect a laboratory accident such as a fire or fire during an
experiment. This will provide help and quick response to the said accident.
6
Pablo Borbon
Tel Nos.: (+63 43) 980-0385; loc 1127
F. Discussion/Conclusion
The purpose of the virtual laboratory exercise was to enable students to see in greater
detail the components of cells and microorganisms that are invisible to the unaided eye. Also,
appropriate lens cleaning must be carried out to prevent the buildup of dust particles, which will
make using the microscope and seeing the specimen uncomfortable. In biomedical science classes
as well as in research and diagnostic labs, the light microscope is a particularly effective tool for
comprehending the structure and function of tissues. Learners are now able to observe cells,
bacteria, and a variety of other objects that are too small to be seen without a microscope.
7
Pablo Borbon
Tel Nos.: (+63 43) 980-0385; loc 1127
Laboratory Activity No 2:
Streak Plate Method
A. Introduction
8
Pablo Borbon
Tel Nos.: (+63 43) 980-0385; loc 1127
The streak plate method in the field of microbiology is crucial as it has the purpose to
collect isolated colonies from a single area with increasing dilution to serve as an inoculum plate,
based from American Society for Microbiology. With this technique, biologists allow
microorganisms such bacteria and fungi to grow on semi-solid surface with agar, a culture media
containing nutrients to produce various colonies. Also, this method can help scientists to
determine the organism with the colonies. The objective of this activity is to know how to
conduct the stated method, and to observe how microorganisms produce different colonies.
Lastly, the main objective of this activity is to know the importance of streak plate method.
B. Materials
 Petri dishes
 Nutrient agar
 Inoculating loop
 Incubator
 Bunsen burner
 Sterile pipettes
 Alcohol or disinfectant
C. Procedure
 Sterilize all the equipment or materials instruments, flasks and media that are required for
the streaking procedure.
 Prevent the spread of microorganism by cleaning your work area using a disinfectant to
minimize any contamination.
 Carefully set up the Bunsen burner in your work area.
9
Pablo Borbon
Tel Nos.: (+63 43) 980-0385; loc 1127
 Wash hands with an antiseptic solution before handling any microbial solution to avoid
any infections.
 Use inoculating loop to pick up the sample. You can use either a metal loop or disposable
plastic loops.
 Sterilize the loop by heating it in

the Bunsen burner until it is red
hot if you are using a metal loop and allow it to cool. Make sure to do this process before
and after use of the loop.
 Pick
an
isolated colony from the agar plate culture and spread it over the first quadrant
(approximately 1/4 of the plate) using close parallel streaks.
10
Pablo Borbon
Tel Nos.: (+63 43) 980-0385; loc 1127
 Streak the other three quadrants by a similar method.
 Close the lid of the plate after streaking, and store the dish upside down. Invert the plate
and incubate at 37°C for 24 hr.
 Observe
D. Guide Questions
1. Why is flame-sterilizing the inoculating loop needle before and after each use
necessary?
Besides on streaking, one of the repetitive processes is flame-sterilizing the
inoculating loop needle. The flame-sterilizing is done before and after use of inoculating
loop needle to make sure that there is no presence of other materials in the tool. If the
inoculating loop needle is not been sterilized, the objective of the experiment won’t be
met. Additionally, if the needle is not sterile, there are possibilities that other bacteria
would spread on the plate causing it not to produce discrete colonies. Also, without
colonies, you won’t have an accurate vision and you won’t identify the organism on the
11
Pablo Borbon
Tel Nos.: (+63 43) 980-0385; loc 1127
particular material. Therefore, it is important to sterilize the inoculating loop needle

before and after use using flame-sterilizing tool, Bunsen burner.
2. What is the advantage of the four-quadrant streaking method?

The four-quadrant streaking method is an improved version of the streak plate
method that offers several advantages. One of the main advantages of the four-quadrant
streaking method is that it allows for the isolation of a greater number of individual
bacterial colonies. This is because the sample is streaked in a cross pattern, creating a
gradient of decreasing bacterial density, which helps to separate individual cells from
each other. Additionally, the four-quadrant streaking method is more efficient in
obtaining pure bacterial cultures, as it reduces the likelihood of contamination by other
microorganisms. This is because the first quadrant is used to dilute the sample, while the
subsequent quadrants are used to isolate individual bacterial colonies. Overall, the four-
quadrant streaking method is a valuable tool in microbiology, as it allows for the efficient
and accurate isolation of pure bacterial cultures.
3. What was the purpose of incubating the inoculated plates after the set-up?
Bacteria growth is usually performed under temperature conditions. Petri plates
are filled with agar, which feeds bacteria that are inoculated on the surface. Under the
proper conditions (usually 37 degrees Celsius), the bacteria will consume the agar as
food and grow into colonies called colony forming units. As streaking method was done
in various quadrants, the plate will now be placed in an incubator that has been set to a
temperature of 37°C. In light of this, incubating is one of the essential requirements of
culture media. Thus, the main purpose of incubating or maintaining the inoculation plate
at the right temperature is to promote the visibility growth of bacterial colonies. This is
for subsequent microbiology study as a crucial component of the investigation.
E. Discussions and Conclusions

In conclusion, with this method the bacterial load is gradually diluted as streaking
progresses over the surface of the agar medium at the end, only a small number of bacteria will be
inoculated, resulting in wellisolated colonies in the final streaks. Hence, using this technique, the
bacteria were mechanically separated from a mixed population of either the same or different
species. If the specimen had a single species, the same types of colonies were evident in the
terminal streaks after inoculation. However, if the specimen contained multiple species, different
types of colonies might have been visible.
12
Pablo Borbon
Tel Nos.: (+63 43) 980-0385; loc 1127
References
Katz, D. S. (2008). The Streak Plate Protocol. American Society for Microbiology.
https://asm.org/ASM/media/Protocol-Images/The-Streak-Plate-Protocol.pdf?ext=.pdf
A. (2022). Streak Plate Technique. BYJUS. https://byjus.com/neet/streak-plate

technique/#:~:text=Streak%20Plate%20Method%20Principle
Atlas, R. M., & Parks, L. C. (2015). Handbook of microbiological media. CRC Press.
13
Pablo Borbon
Tel Nos.: (+63 43) 980-0385; loc 1127
https://books.google.com.ph/books?
hl=en&lr=&id=NpbLBQAAQBAJ&oi=fnd&pg=PP1&dq=Atlas,+R.+M.,+%26+Parks,+L.+C.
+(2015).+Handbook+of+microbiological+media.
+CRC+Press&ots=QBo4vY_rAA&sig=DGlBarcurfClPILBHVpyx0NbFhw&redir_esc=y#v=one
page&q&f=false Bacterial Growth. (n.d.). Revolutionary Science.
https://www.revsci.com/pages/bacterial-growt
https://www.leica-microsystems.com/science-lab/applied/what-does-300001-magnification-
reallymean/#:~:text=Magnification%20is%20the%20ability%20of,object%20with%20the%20un
aided%20eye.
https://www.azooptics.com/Article.aspx?ArticleID=2339#:~:text=The%20microscope%20lens
%20functions %20like,ocular%20lens%20near%20the%20observer.
https://www.thoughtco.com/important-lab-safety-rules-608156
About the Author
This lab report was written by Kelvym A. Macalintal, a student of Bachelor of Nursing in
Batangas State University TNEU. With a strong interest in science and technology, He has
always been passionate about exploring new topics and finding solutions to problems.
14
Pablo Borbon
Tel Nos.: (+63 43) 980-0385; loc 1127
15

Laboratory Report in Microbiology and Parasitology

Uploaded by

Document Information

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Laboratory Report in Microbiology and Parasitology

Uploaded by

Copyright:

Available Formats

Republic of the Philippines

BATANGAS STATE UNIVERSITY

D. Data and Analysis

Actual Sample from

Description/ Use of • Acid-fast organisms • Endospores • A Gram stain is

 Sterilize the loop by heating it in

 Streak the other three quadrants by a similar method.

particular material. Therefore, it is important to sterilize the inoculating loop needle

2. What is the advantage of the four-quadrant streaking method?

E. Discussions and Conclusions

A. (2022). Streak Plate Technique. BYJUS. https://byjus.com/neet/streak-plate

About the Author

You might also like