MC3 MICROBIOLOGY AND PARASITOLOGY

BSN 1 – E

Performance Task 1 (Lab):

Virtual Laboratory Report on Examination of Microorganisms
in Living State

GROUP 3

Members:

❖ Andrada, Marko Polo Carmelo

❖ Dreo, Zyla Glianne
❖ Dioso, Krizel Ann
❖ Geraneo, Kayah Sharley
❖ Gerochi, Janine Jertrude
❖ Jaleco, Eloisa Mae
❖ Labanero, Khessie Mae
I. Title of experiment

Examination of Microorganisms in Living State

II. Short introduction

Microorganisms or microbes are living things, are ones that are called to be
'invisible/impossible to be seen with the naked eye.' And prolly existed first before humans took
over the world. Leeuwenhoek, the first person who discovered the microorganisms together with
his co-scientist Hooke; he described microorganisms as dierkens or in English, small animals
that can only be seen under a microscope.

Microorganisms can be classified as bacteria, fungi, archaea and protists, or even

viruses. These microorganisms are widespread in nature and beneficial to life, however there
are some that can cause a threat and harm to one's health— these microorganisms are the
pathogenic or bad bacteria and viruses which then causes illnesses, food poisoning, STDs and
other transmitted diseases.

III. Materials/specimen used

HOW TO PREPARE A WET MOUNT

Gloves Cover slip

Googles Deionized water
liquid Inoculating loop
Colony on a Petri plate Bunsen burner
Slide

HANGING DROP PREPARATION TO DEMONSTRATE BACTERIAL MOTILITY

Young broth culture Loop

Slide with adhesive or paraffin ring/ circular Cover slip
depression Bunsen burner
Paraffin wax microscope
IV. Methods/procedure

Demonstration on how to do a wet mount when the starting material is a liquid and then when
the starting material is coming from a colony on a Petri plate.

Starting material is a liquid

1. Take a slide from the box.

2. Get a cover slip (cover slips are squares of either glass or plastic and they are
separated by pieces of tissue paper).

3. Take a drop of the material and put it in the center of the slide.

4. Take the cover slip and hold it at a 45 degree angle touching the slide and gently drop
it.

Starting material is a colony on a Petri plate

1. Take a slide

2. Put a drop of deionized water in the center of the slide.

3. Use aseptic technique using the inoculating loop.

4. Sterilize the inoculating loop by putting the loop in the flame of the Bunsen burner.

5. Remove the lid from the plate if there’s an area on the plate where you can touch the
loop to cool it beforehand.

6. Touch the surface of a colony and immediately put the lid back on the plate.

7. Put the loop in the drop of water.

8. Spread it around about the size of a dime.

9. Sterilize the loop before you put it on the bench.

10. Turn off the Bunsen burner.

11. Take a cover slip, hold it at a 45 degree angle and gently drop it on the sample.
Hanging drop preparation to demonstrate bacterial motility.

1. Get a slide with a paraffin ring, adhesive tape ring, and circular concavity.

2. Take cover slips and place it in the upper part of each slides.

3. Place a loopful of broth on the cover slips.

4. Sterilize the loop in flame.

5. Apply small amounts of paraffin wax around the ring or concavity to make it sticky.

6. Invert the slide with its sticky side facing downwards.

7. Gently press against the cover slip and revert it quickly.

8. Now the drop hangs on the cover slip and ready to be observe under the microscope.

V. Results of experiments

After the experiment on how to prepare wet mount, wet mount can be used when you
need or want to visualize a living microorganism. For example if you wanted to determine if an
organism is motile (able to move) or not. In the experiment hanging drop preparation in which a
drop of medium containing the organisms is placed in the microscope slide, often is used to
observe the motility of the bacteria. The result shows that if it is observed in the microscope
under dry low power objective (10x) lens, only the edge of the drop is seen but if it is observed
under 40x objective lens, active motile bacilli can be determined.

VI. Discussion

The wet mount and hanging drop preparations are both used to observe living microbes
and organisms. With the help of these two preparations, the motility of the organism can be
seen, as well as the original shape and size of the organism.

Wet mount increases the transparency of the specimen and makes it easier to stain. The
use of a wet mount slide flattens the specimen, making it simpler to view and observe. By using
this type of method, the size and shape of individual organisms can be seen as well as the
characteristic arrangement or groupings of cells. The wet mount method is also used to
determine whether or not an organism is motile (able to move). In the video, the materials that
were needed in order to perform such method are gloves, goggles, liquid, and colony on a Petri
plate, slide, cover slip, deionized water, inoculating loop, and Bunsen burner. In a wet mount,
the specimen is suspended in a drop of liquid (typically water) between the slide and the cover
glass. The water's refractive index enhances image quality while also supporting the specimen.
Wet mounts, unlike permanently fixed slides, cannot be stored for long periods of time because
the water evaporates.

Hanging drop, on the other hand, is a significant technique for studying bacterial motility
as well as the form, size, and arrangement of bacteria. It does not distort the form or
arrangement of the cells. Furthermore, the hanging drop method allows for a better view of
bacterial motility than the wet mount method. The wet mount dries out fast when exposed to the
heat of the microscope light; it is simpler to perform than the wet mount, but it is useful for
short-term observation only. Materials that were needed in the video experiment were the
following: young broth culture, slide with adhesive or paraffin, ring or circular depression,
paraffin wax, loop, cover slip, Bunsen burner and the microscope. By performing the said
method, results showed that when a specific microbe is studied in a microscope with a dry low
power objective 10x lens, only the edge of the drop can be seen. Meanwhile, active motile bacilli
can be seen under a 40x objective lens.

The shape and size of bacteria can be seen in wet mount, however motility may be
hampered since the suspension is pressed between the slide and the cover slip. As a result, a
hanging drop preparation or motility test is used to observe the movement of bacteria in addition
to their shape and size.

VII. Conclusion

All in all in this task we are able to identify the motility and the size of a cell with the
proper way of using aseptic technique.

VIII. Short reflection of the experiment

As nursing students, we believe that these experiments will assist us in learning,

understanding, and eventually practicing various microscopy techniques to accomplish the
required observation results given the specimen and particular areas of the specimen. Wet
mount and hanging drop microscopy are two microscopy techniques that may be used to
examine a wide range of live microorganisms. Although we cannot see or experience the
microorganisms in the videos, we are grateful that, despite the pandemic, we have the
opportunity to study and learn about microbiology and parasitology.
IX. List of references

Microbiology, M. (2015, August 25). How to Prepare a Wet Mount - MCCC Microbiology.

YouTube. https://www.youtube.com/watch?v=N1780tJTk90&feature=youtu.be

Rao, S. (2010, June 11). Hanging drop for bacterial motility. YouTube.
https://www.youtube.com/watch?v=ujzSmsmg7ok&feature=youtu.be

Microbehunter Microscopy. (2021, July 17). Making a wet mount microscope slide |
Microbehunter Microscopy. MicrobeHunter.Com.
https://www.microbehunter.com/making-a-wet-mount-microscope-slide/?fbclid=IwAR1c9
8gTVzt_x06N0_iKZDTVKWCqkDTdYzxbZJqu5Zxcyusdiqq5pg9ySqg

Tankeshwar, A. (2021, June 17). Hanging Drop Method for Bacterial Motility. Microbe Online.
https://microbeonline.com/procedure-hanging-drop-method-test-bacterial-motility/?fbclid
=IwAR2N755XdJY8yeY3gpleCq4R1mISeuhzJEJvdJ5QA-SpzAKKUe3aYCjIvoY

N, S. (2020, December 26). What is Hanging Drop Method? Definition, Procedure &
Applications. Biology Reader.
https://biologyreader.com/hanging-drop-method.html?fbclid=IwAR0J3XB8U_KBHy4EU4
BZTtdlggpX1dFNIhKMxVHgFSgGYWUq9QcEBv5ASlQ#:%7E:text=using%20this%20m
ethod.-,Advantages,than%20the%20wet%20mount%20method

Libretexts. (2021, January 3). 1.2A Types of Microorganisms. Biology LibreTexts.

https://bio.libretexts.org/Bookshelves/Microbiology/Book%3A_Microbiology_(Boundless)/
1%3A_Introduction_to_Microbiology/1.2%3A_Microbes_and_the_World/1.2A_Types_of_
Microorganisms?fbclid=IwAR2zVt_MJvLZtXY39OgEG2QSMWvLQuC4Ok9v1pNyBZe2v
-Gt4_yNMyVov6s
Members who contributed:

❖ Andrada, Marko Polo Carmelo

❖ Dreo, Zyla Glianne
❖ Dioso, Krizel Ann
❖ Geraneo, Kayah Sharley
❖ Gerochi, Janine Jertrude
❖ Jaleco, Eloisa Mae
❖ Labanero, Khessie Mae