THE ENTIRE SEMESTER'S SET OF EXPERIMENTS

Laboratory Report in Microbiology and Parasitology - MC102L

Presented to

In Partial Fulfillment

Of the Requirements for

MC 102L MICROBIOLOGY AND PARASITOLOGY ( LABORATORY )

May 8, 2023
 TABLE OF CONTENTS

Page

TITLE PAGE ……………………………………………………………………………………… 1

TABLE OF CONTENTS …………………………………………………………………………. 2

LIST OF TABLES ………………………………………………………………………………… 3

LABORATORY ACTIVITY 1: ( Manipulating a Microscope )

I. Introduction ……………………………………………………………………………….... 4

II. Materials ………………………………………………………………………………......... 4

III. Procedure …………………………………………………………………………………... 5

IV. Data Analysis .…………………....………………………………………………………… 8

V. Guide Questions .………………….………………………………………………………… 10

VI. Discussions and Conclusions ………………………………………………………………. 13

LABORATORY ACTIVITY 2: ( Streak Plate Method )

I. Introduction ……………………………………………………………………………….... 15

II. Materials ………………………………………………………………………………......... 15

III. Procedure .………………………………………………………………………………….... 16

IV. Guide Questions .………………….………………………………………………………… 18

V. Discussions and Conclusions ….……………………………………………………………. 19

REFERENCES ……………………………………………………………………………………… 21

ABOUT THE AUTHOR ……………………………………………………………………………. 23

LIST OF TABLES

Table No.1 Title

Page

1 Procedures Rendered During the Experiment 5

2 Acid Fast Mix 8

3 Endospore Stain 9

4 Gram Stain Mix 10

5 Procedures Conducted During the Activity 16

 LABORATORY ACTIVITY NO. 1:

MANIPULATING A MICROSCOPE

I. INTRODUCTION

Microscopes are optical equipment that allows us to see the microbiome. Without a

microscope, there would be nothing to do in a microbiology laboratory since the objects of our interest

(bacteria, fungus, and other single-celled animals) are too tiny to view. Due to limited access to the

microscope, the students completed the exercise using a virtual microscope. Students may learn

vocabulary, usage, and care using a fully working virtual microscope. Its goals are to identify the

fundamental components of a standard microscope, describe how to utilize lens power and eyepiece

powers, calculate the magnification of a microscope depending on the selected lens, and discuss how to

care for and operate a typical microscope.

II. MATERIALS

1. Laptop

2. Phone

3. Ballpen

4. Yellow Paper

5. YOUTUBE LINK: https://youtu.be/3ZFW_nbV7RM

6. SIMULATION LINK: https://www.ncbionetwork.org/educational-

resources/elearning/interactive-elearning-tools/virtual-microscope

III. PROCEDURE

Table 1

Procedures Rendered During the Experiment

14 steps in inspecting the bacterial Documentation

slide with a virtual microscope.
Step 1: Refer to the link:

https://www.ncbionetwork.org/educatio

nal-resources/elearning/interactive-

elearning-tools/virtual-microscope

Step 2: After the screen loads, other

tabs will appear, but pick the explorer

tab to utilize the microscope.

Step 3: Tap the only thing with a

clickable question which is the slide

box.

Step 4: After clicking the slide box,

possible slides will be shown.

Step 5: Select the bacterial slide and

inspect the three created slides using a

microscope to do the task successfully.

Step 6: Adjust the slider bars starting

with the coarse focus; the more you

move the coarse focus, the clearer it

becomes.

Step 7: Alter the fine focus to improve

the image's focus quality. You can

change the emphasis on each slide until

it appears as nice as possible.

Step 8: Adjust the quantity of light

flowing through the condenser by

moving the light adjust. You can

change the brightness in each slide till

it appears as well as it can.

Step 9: Going on to the next

magnification, 10x, you've zoomed in

slightly and have the option of

dragging the circle that specimen

around.

Step 10: Heading on to the next

magnification, 40x, you are zoomed in

and have the option of dragging the

circle that specimen around.

Step 11: When you reach the 100x

magnification, apply immersion oil to

increase resolving power. You can drag

the circle that specimen around since

you're extremely zoomed in.

Step 12: Clean the lens and slide with

the lens paper after using the 100x lens

with immersion oil.

Step 13: While repositioning the

microscope, begin at 100x and

gradually decrease the magnification.

 Returning it to 4x may harm the lens,

slide, or microscope.

Step 14: To return to the main screen,

click the remove slide and then the

main arrow.

IV. DATA AND ANALYSIS

BACTERIAL SLIDES

TABLE 2

Acid Fast Mix

Description / Use Of The Mycobacterium smegmatis

Sample

Microscope View On
Different Focus And
Magnification

Sample From Internet

Reference Of The Sample https://open.maricopa.edu/redmountainmicro/chapter/acid-fast-stain/

From Internet

TABLE 3

Endospore Stain

Description / Use Of The Bacillus subtilis at 2 weeks old

Sample
 Microscope View On
Different Focus And
Magnification

Sample From Internet

Reference Of The Sample https://microbeonline.com/wp-content/uploads/2015/05/Spore-Stain.jpg

From Internet

TABLE 4

Gram Stain Mix

Description / Use Of The Bacillus subtilis

Sample

Microscope View On
Different Focus And
Magnification

Sample From Internet

Reference Of The Sample https://www.austincc.edu/microbugz/gram_stain.php

From Internet

V. GUIDE QUESTIONS

1. What is magnification? What is its importance in microscopy?

 The capacity of a microscope to generate a picture of an item at a scale bigger (or even

smaller) than its actual size is referred to as magnification. The process of growing the perceived size

of anything, rather than the actual size, is known as magnification. A determined number quantifies its

magnification. When this number is less than one, it indicates a size reduction, also known as

magnification or de-magnification (Brittanica,2011). It is crucial in microscopy because the picture

allows the user to see more details of an item than when studying the object with the naked eye.

Multiples of magnification, such as 2x, 4x, and 10x, indicate that the item has been expanded to twice

its original size, four times its original size, or ten times its original size, respectively.

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.

To be capable of operating something, one must be thoroughly familiar with its many operable

pieces. As a result, based on the experiment, below are some of the most commonly utilized parts.

The objective lenses give additional magnification and are adjustable. Objective lenses are

among the most significant components of an optical microscope since they are responsible for primary

image generation and play a critical role in defining the quality of pictures produced by the

microscope. Users can turn a knob or move the binocular lenses in the activity, simulating the

modifications of the natural lens in the eyes that allow the user to see items at varying distances. A

microscope typically has three or four objective lenses. The most popular are 4X (the shortest lens),

10X, 40X, and 100X (the longest lens), all used in the activity to examine the bacterial slides. The

higher magnification objectives (beginning with 40x) are spring loaded. If a spring-loaded objective

lens collides with a slide, it will retract, protecting both the lens and the slide (AmScope, 2020). The

picture of an item may be enlarged and examined in depth using the microscope's lenses.

According to the experiment results, the illuminator is the second most commonly utilized

component. The illuminator's light brightness may be adjusted to fit the preferences and what the user

is seeing. It delivers even, high-intensity light at the field aperture, allowing light to flow through the

condenser to the specimen. It was employed in the exercise because we could alter the illumination to

our liking, which let us see the germs more clearly. It improves viewing since there is no replacement

for a microscope with built-in high-quality illumination that delivers natural color for studying objects

and specimens.

The final part is the eyepiece, which is used to enlarge the picture generated by the objective

lens. It is situated above the stage and is used to examine the specimen under the microscope. These
lenses, sometimes known as ocular lenses, are normally 10X, although they are also available in 5X,

15X, and 20X sizes. The user view the specimen through the eyepiece lens when the user gaze through

it. The interocular distance may be adjusted so the user can look through the microscope with both eyes

open. It is employed in the activity to amplify the principal picture generated by the objective, allowing

the eye to exploit the full resolution capacity of the objective. It also has a diopter adjustment, allowing

the user to tailor the eyepiece to their vision. Overall, the eyepiece is a crucial component of this

activity since it is utilized to enlarge the picture generated by the objective lens, allowing the user to

examine the specimen in more detail.

3. When switching lenses in a microscope, why is it important to move through lenses in

order?

When switching from low to high power on a microscope, the high-power objective lens

travels directly over the specimen while the low-power objective lens rotates away. Users should go

through the lenses gently to improve magnification while avoiding damage to the microscope. This

modification affects a specimen's magnification, light intensity, field of vision area, depth of field,

working distance, and resolution (Murphy,2018). If the lenses are of high quality, the image should

remain focused. When the user increases the power, the field of view narrows. On low power, the user

will see more of an object. The lowest power objective has the greatest depth of concentration. The

depth of focus is diminished each time the user switches to a higher power setting. As a result, with

increased magnification, a smaller portion of the specimen is in focus.

As the magnification is increased by switching to a higher power lens, the working distance

drops and the user sees a much smaller slice of the specimen. Furthermore, because the lenses on the

microscope are designed to be parfocal, the user should not have to make any substantial modifications

while switching lenses once focussed on an item. This implies that if the user saw something in focus

with the low power objective, it should be virtually in focus while switching to the high power

objective, and vice versa. Consequently, regardless of the lens used to examine the object, the ideal

approach is to initially set the working distance with a lower power lens then adjust it to proper focus

using the coarse focus knob.

4. For you, what is the most important laboratory safety rule? Justify your answer.

The most crucial laboratory safety guideline for me was closing the lab door. Most people

consider lab doors merely workplace entrances and exits, but they are also critical for life safety and
hazard control. While moving between rooms regularly, it is normal to prop open lab doors. This

approach is more convenient, but it jeopardizes the safety of the laboratory and its surroundings.

Closed lab doors aid in maintaining negative air pressure. The structures are built with negative

directional air flow from the hallway to the lab areas in mind. The capacity of the building to

accomplish this is jeopardized when the doors are left open. This is also a crucial part of managing

campus fire safety and biosecurity.

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)

If I had to design one lab instrument, it would be goggles with lenses that could be adjusted to

fit your glass eye level. It wasn't discussed much how awkward it is to wear glasses within the goggles;

it's nearly impossible to experiment without goggles, but getting those goggles to function with

prescription glasses may be difficult. They have a bad habit of failing to form a good seal, fogging, and

being uncomfortable. That's why I designed these goggles with changeable lenses to ensure that

everyone can see well throughout lab tests. Instead of solid, one-of-a-kind lenses, these glasses contain

a hard lens in front and a softer, more flexible plastic sheet in back. A layer of viscous liquid with a

high refractive index exists between them (light-bending ability). You may custom fit your spectacles

to yourself by pumping more or less of this liquid between the layers; no pricey opticians or lens-

grinding required.

VI. DISCUSSIONS AND CONCLUSIONS

This practice aids the student's knowledge of the main concept areas in the microbiology and

parasitology labs and the development of an investigative and analytic thinking approach to clinical

circumstances. As digital biology advances, this technology will surely be a helpful tool for enhancing

uniformity, dependability, and validity in the medical course. This lab material will assist students in

better understanding the role of information technology in health care choices and patient education.

The advantages of virtual microscopes are undeniably important for medical students, pathologists, lab

workers, and researchers who wish to become skilled in using this innovative technology. Adding

virtual microscopy to conventional microscopy improves knowledge of the subject compared to the

usage of conventional microscopy and virtual microscopy alone.

 LABORATORY ACTIVITY NO. 2:

STREAK PLATE METHOD

I. INTRODUCTION

The streak plate method is a quick and effective qualitative isolation method. The streak plate

technique is used to grow bacteria on a growth media surface in order to isolate and sample individual

bacterial colonies. The modern streak plate method evolved from Robert Koch’s and other

microbiologists’ efforts to obtain pure cultures of bacteria in order to study them. In this activity,

students shall be able to observe and conduct the streak plate method through virtual simulation.

Considering the objectives of obtaining a pure culture of bacteria from a mixed culture; obtain well

isolated colonies and propagate bacteria.

II. MATERIALS
1. Inoculation Loop

2. Bunsen Burner

3. Agar Plate
 4. Petri Dish

5. Timer

6. Incubator

1.
2. 3.

4. 5. 6.

III. PROCEDURES

TABLE 5

Procedures Conducted During the Activity

Procedures Photo Documentation

STEP 1: Click the simulation link. The student will

be directed to the website.

STEP 2: For better understanding about the

experiment, proceed to the “Description.” It

contains the purpose of the experiment, the method,

and the interpretation. Click “x” on the upper right

to go back to the Main Menu.

STEP 3: Click “Steps”to review the procedure.

Click “x” on the upper right to go back to the Main

Menu.
STEP 4: Select “Start” to commence the

experiment.

STEP 5: Drag the inoculation loop to the Bunsen

burner. Sterilize it over the flame until the wire

turns red and then let it cool afterwards.

STEP 6: After sterilization, pick one individual

colony from the agar plate.

STEP 7: Spread the colony back and forth on the

first quadrant of the plate using close, parallel

streaks.

STEP 8: Sterilize the inoculating loop once again

over the flame. Allow it to cool afterwards.

STEP 9: Again, streak into the second quadrant

back and forth.

STEP 10: For the third time, sterilize the loop over

the Bunsen burner, allowing it to cool later on.

STEP 11: Streak to another quadrant using the

same technique.

STEP 12: Sterilize the inoculation loop until hot red

and let it cool.

STEP 13: Proceed to streaking on the last

quadrant.

STEP 14: Place the plate in an incubator with a

37°C temperature for 24 hours.

STEP 15: Closely observe the result and gather the

important data.

IV. GUIDE QUESTIONS

1. Why is flame-sterilizing the inoculating loop needle before and after each use

necessary?

In the streak plate test, the inoculating loop needle must be flame-sterilized both before and

after each usage. First, to avoid contamination, the surface of the inoculating loop is flame sterilized to

get rid of any potential pollutants. This makes it more likely that the bacteria under test are the only

ones in the sample and that any observed growth is due to those same germs. Second, cross-

contamination must be prevented. Cross-contamination between samples is another thing that flame

sterilization helps to avoid. Bacteria from one sample may be transferred to the following sample if the

inoculating loop is not sterilized after each use. This could produce unreliable results. The need to

provide accurate outcomes is equally essential. To separate specific bacteria, utilize the streak plate

test. Individual bacterial colonies can be isolated using the streak plate test for future research.

Obtaining pure cultures that are uncontaminated by other organisms is necessary for accurate results.

The danger of false-positive or false-negative results is decreased by sterilizing the inoculating loop

before and after each usage, which guarantees that only the target bacteria are present in the sample.

Overall, a crucial step in the streak plate test that helps guarantee precise and trustworthy results is

flame-sterilizing the inoculating loop before and after each usage.

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

The four-quadrant streaking method’s benefits include the ability to separate specific colonies

from the mixed culture, which is important for colony isolation. The amount of germs scattered onto

the plate in each quadrant is reduced by dividing the plate into four quadrants and using a sterile loop
to streak the bacteria, increasing the possibility of single colonies developing. Next is colony

morphology, which can be observed using the four-quadrant streaking method. Colony morphology

can reveal important details about the identity and traits of the bacteria. Last but not least is the ease of

interpretation. The technique offers a clear and organized way of streaking bacteria, making it simpler

to interpret the findings and recognize the colonies of interest. In general, the four-quadrant streaking

method is a straightforward and efficient way to isolate bacterial colonies. It can also reveal important

details about the properties of the bacteria being studied.

3. What was the purpose of incubating the inoculated plates after the set-up?

The purpose of incubating the inoculated plates after the setup of a streak plate test is to

provide optimal growth conditions for the microorganisms that were streaked onto the plate. The

microorganisms can grow and form colonies that can be seen during the incubation process. These

colonies can then be examined and studied for purposes of identification and characterization. The

streak plate test requires the researcher or students to monitor the growth and morphology of the

microorganisms; hence, incubation is a crucial step. Many details about the bacteria, including their

metabolic properties, pathogenicity, and antibiotic resistance, can be learned from the colonies’

appearance and development rate.

V. DISCUSSIONS AND CONCLUSIONS

The streaking method is a successful process for isolating an individual specie because cell

density decreases within each quadrant and cells have more space to grow into isolated colonies.

Conducting this activity virtually made us realize that it could be much easier to perform on a face-to-

face set-up, because the instructor could guide us. Procedures must be known beforehand to avoid

delay of the activity; and performing it appropriately which helps in getting the correct results. The

inoculating loop must be flamed before and after each usage to avoid contamination, and cross-

contamination between samples. When streaking the bacteria off with the loop, the agar surface on the

bacteria was rubbed off until the bacteria separated, which aids in the shaping, elevation, pigmentation,

margin, and tex

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AmScope. (2020). Microscope Parts and Functions. Microscope Parts & Functions – AmScope,

3(5), 1-2. https://amscope.com/pages/microscope-parts-and-functions/pdf

Aseptic technique. (n.d.). SCIENTIST CINDY. https://www.scientistcindy.com/aseptic-technique.html

Britannica, T. (2011). Magnification. Encyclopedia Britannica.,

https://www.britannica.com/technology/magnification

Gram Stain. (n.d.). Welcome to Microbug. Gram

Stain., https://www.austincc.edu/microbugz/gram_stain.php

I. (n.d.). Incubating and viewing plates. https://practicalbiology.org/standard-techniques/incubating-

and-viewing-plates

Malachite green staining. (n.d.). Endospore staining. Endospore staining procedure.,

https://microbiologie-clinique.com/endospore-staining-malachite-green.html

Murphy, E. (2018). SCIENCING. What Happens When You Go From Low Power to High Power on

a Microscope?, 23(7), 29-31. https://sciencing.com/oil-immersion-lens-needed-bacteria-

19559/xml/jats

N, S. (2021, December 13). Quadrant Streaking Method – Meaning, Principle, Diagram & Procedure.

Biology Reader. https://biologyreader.com/quadrant-streaking-method.html

Raymond, J., PhD. (2022). Pressbooks. ACID-FAST

STAIN. https://open.maricopa.edu/redmountainmicro/chapter/acid-fast-stain/

Streak Plate Test. https://learn.chm.msu.edu/vibl/vibl/StreakPlate/streak_plate_HTML5Canvas.html

Tankeshwar, A. (2022). Streak Plate Method: Principle, Procedure, Uses.

https://microbeonline.com/streak-plate-method-principle-purpose-procedure-results/

Vlab.amrita.edu,. (2011). Streak Plate Method. https://vlab.amrita.edu/index.php?

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