Laboratory Report No.

3
Microbial Size Determination Using Compound Microscopy
submitted by: Nicky Joy Ramasta
BSEd Science III
Eastern Samar State University-Salcedo Campus
[email protected]

submitted to: Rica C. Elacion

Professor
Eastern Samar State University-Salcedo Campus
[email protected]

Date Submitted: January 24, 2024

Microscopic life forms, which are often Fig. 1. Calculation for the actual size (field of
too small and invisible to the naked eye, come in view measurement).
a range of shapes and sizes. They play a role, in
processes, human health and various industrial Field of view
applications. By studying these organisms under Cell Size
Fit number
a microscope, gaining insights regarding the =
physiology, behavior and potential uses become Fig. 2. Calculation for the actual cell size.
possible. One important aspect of understanding
microorganisms is determining the sizes since This laboratory experiment focuses on
this information is crucial for comprehending measuring the sizes of microorganisms using a
the physiology, taxonomy and ecological compound microscope. The experiment involves
functions of these microorganisms. preparing samples of different microbes,
applying staining techniques to improve contrast
In research the use of a compound during analysis for measuring cell dimensions
microscope is fundamental as it allows the (2).
visualization of microorganisms with resolution.
Accurate measurements of sizes are vital across Understanding microorganism sizes is
scientific fields like microbiology, medicine and not crucial for research but, holds practical
environmental science. Precise size implications. In fields like medicine,
determination helps identify species classify understanding the dimensions of cells is
them appropriately and establishes a foundation essential to determine the dosage of antibiotics
for studying the diversity within communities and to comprehend how infections work. In
science, measuring the size of microorganisms
plays a role in evaluating their functional
contributions, within ecosystems (3).

(1).
 Examine and select a variety of prepared slides
Fig. 3. Illustration of compound microscope. with different specimens. Ensure that specimens
The objectives of this experiment are are well-positioned on slides, and coverslips are
twofold: first, to accurately measure the sizes of securely placed.
selected microorganisms, and second, to Begin with the lowest magnification
appreciate the significance of microbial size objective (4x) for initial specimen location.
determination in broader scientific contexts. The Progressively switch to higher magnifications
experiment employs a compound microscope, a (10x, 40x, and 100x) for detailed observations.
tool that has been pivotal in advancing our Place a prepared slide on the microscope stage
understanding of the microbial world since the and secure it with the stage clips. Use the coarse
early days of microbiology. focus knob to bring the specimen into
approximate focus. Adjust the illumination
The observations and measurements intensity to enhance contrast. Switch to higher
gathered from this experiment provide valuable magnification objectives (10x and above).
data for understanding the morphological Utilize the fine focus knob for precise
diversity of microorganisms. This information adjustments to achieve optimal clarity. Focus
contributes not only to the fundamental incrementally to observe fine details of the
knowledge of microbial biology but lays the specimen.
groundwork for further research and applications
in diverse scientific disciplines. Explore the concept of depth of field by
focusing on different planes within a specimen.
In the subsequent sections of this report, Observe how adjusting the focus impacts the
materials and methods employed in the clarity of structures at varying depths. Practice
experiment, present the results of size focusing on specimens using the 4x and 10x
measurements, and engage in a comprehensive objectives. Experiment with different specimens
discussion of the findings. to understand variations in focusing
requirements. Participants progress to higher
Materials magnifications (40x and 100x). Focus on finer
details and structures within the specimens.
 Text book (Reference)
 Compound light microscope Record observations in lab notebooks.
 Microscope slides Document challenges faced, adjustments made,
 Coverslips and notable findings during the focusing
 Medicine dropper exercises. Verify the accuracy of measurements
 Dissecting needle (needle probe) using the calibrated eyepiece reticle against a
 Specimen examples for Lichen and stage micrometer. Adjust the eyepiece reticle
Fungi calibration if necessary.
 Iodine stain
 Micrometer Turn off the microscope and the
illumination source. Clean the microscope lenses
Procedure using lens cleaning solution and lens paper.
Ensure slides and coverslips are properly stored.
Ensure the compound microscope is Facilitate a discussion session to review
placed on a stable and level surface. Adjust the observations, challenges, and key learnings.
interpupillary distance and eyepiece diopter Discuss the importance of proper focusing in
settings. Turn on the microscope's illumination microscopy and its impact on scientific
source and set it to an appropriate intensity. observations.
Calibrate the eyepiece reticle using a calibration
slide with a standard micrometer scale. Adjust This detailed procedure ensures that
the eyepiece to ensure accurate measurements. participants gain hands-on experience with the
measuring techniques using a compound
microscope. It provides a structured approach Fig. 4. Appearance of a. Lichen and b. Fungi
for participants to explore various under Compound microscope.
magnifications, practice measuring different Table 1
specimens, and understand the nuances of
achieving optimal clarity. The discussion session Lichen
encourages reflection on the importance of Appro.
measurement in microscopic observations. Appearance Actual
Cell
Mag. under Size
Measure-
microscope (FOV)
Safety Precautions ment

Compound Microscope is an expensive

and delicate instrument hence, its safety is a 40x 4000μm 4μm
must. Keep the microscope under its dustcover
and in a cabinet when it is not being used.
Handle the microscope with great care. To move
it or carry it, hold it by its arm with one hand
and support its base with the other hand. Store 100x
the microscope with the scanning or lowest 1600μm 32μm
power in place and always start with this
objective when using. To clean the lenses, rub
them gently with the lens paper or high-quality
cotton swab. If the microscope has a substage
lamp, be sure the electric cord does not hang off 400x
the laboratory table where someone might trip 400 μm 40 μm
over it. Do not drag the microscope across the
laboratory table after placing it. Never remove
T. 1. Cell size of Lichen under Compound
parts of the microscope or try to disassemble the
microscope with varying magnification.
eyepiece or objective lenses.
Table 2
Results and Discussion
It was then observed that the field of Agavoideae (Century Plant)
view changes considering its power objective. Appro.
Appearance Actual
Observations revealed the approximate Cell
Mag. under Size
measurement of the microorganisms (Lichens Measure-
microscope (FOV)
and Fungi). The results include the ment
measurements of cells obtained through analysis
and measurement techniques under a
microscope. This data not gives insights into the
physical characteristics of these microorganisms 40x 1000μm 100μm
but also serves as a foundation for further
discussions on size differences comparisons
between different strains and their implications,
for broader scientific knowledge (Fig. 4).

100x 1600μm 160μm

a. Lichen b. Fungi
 include variability in growth conditions,
environmental forces and genetic dissimilarities
across populations. When one can measure and
400x 400 μm 40 μm enumerate such fluctuations, understanding
microbial morphology become better.
Several things account for the varying
T. 2. Cell size of Fungi under Compound sizes of microorganisms under observation.
microscope with varying magnification. These range from differences in genes among
different lines, ecological factors experienced by
The tables show the cell size of lichen organisms during cultivation as well as the effect
and fungi in different magnification. of staining procedures on samples’ sizes. The
Identification of each magnification was based use of dyes in measuring size emphasizes how
on (Table 1) (2).
crucial it is to be cautious when planning
Understanding the sizes of experiments and interpreting microscopy-based
microorganisms is crucial, for unraveling their results.
roles. The size of these organisms plays a role in Understanding environmental roles
how they use resources interact with competitors
among microbial communities requires
and respond to changes in their environment.
understanding of the sizes of microbes. The size
The findings enhance understanding of ecology
and have potential applications in fields like impacts on nutrient acquisition, interactions with
biotechnology and environmental management other microorganisms and the whole ecological
(4). niche occupied by a particular strain. The
deviations in sizes observed reveal potential
To summarize the measurements of adaptive ecological features and mechanisms
microorganism sizes obtained from this employed by microorganisms in their respective
experiment provide insights, into their forms. habitats.
The observed variations highlight the nature of
communities. Moving forward it is important to Knowledge from this experiment is not
address any limitations and conduct studies to only relevant to microbiology but also cuts
refine understanding of microbial sizes and their across different scientific disciplines. Microbial
ecological implications. sizes are important in fields such as medicine,
environmental science, biotechnology etc. This
Generalization study provides basic knowledge about microbial
traits that can be understood in any context.
The revelations from this test give a
critical scrutinizes of the various dimensions This research offers avenues for future
associated with microscopic creatures such as exploration. Including more types of staining
lichen and fungi that were measured using a techniques, examining how specific growth
compound microscope. The measurements given conditions alter the morphology of
therefore lay basis to making assumptions and microorganisms and extending the study to
wider conclusions related to microbiology in cover a wide range of microorganisms may be
particular as a field of science. areas for further investigation. It is important to
consider these issues so as to have better
Different sizes of microorganisms that understanding of microbial sizes.
were observed illustrate the basic diversity of
bacterial and fungal strains within and between Beyond its scientific importance, this
them with a lot of diversity. Factors contributing experiment has an academic worth. The process
to this are not peculiar to species alone but of measuring microbial sizes using a compound
microscope is a useful practical activity for
microbiology students. The integration of theory Appendices
and practice improves learning outcomes and
creates a greater awareness about the intricacies
of microbial life.
In conclusion, the results from this
experiment lead to more detailed knowledge on
microbial sizes and their significance. The
observed variations show how complex
microorganisms are and emphasize the necessity Fig. 5. Viewing the specimen under Compound
for comprehensive investigations that will microscope with a magnification of 40x.
unravel complexities within bacterial
communities. These fundamental facts aid in
current scientific efforts and provide building
blocks for future discoveries in microbiology.

References
Fig. 6. Switching the objective power form low-
1. Experiment to Measure the Size of
power objective to high-power objective.
Microorganisms under Microscope
(With Figure). (2014, February 18).
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size-of-microorganisms-under-
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24, 2024.
2. Measuring the size of Specimens viewed
Fig. 7. Viewing the specimen under Compound
with a Microscope and adding Scale
microscope with a magnification of 100x.
Bars to Photomicrographs. (n.d.). Motic
Microscopes.
https://moticmicroscopes.com/blogs/arti
cles/measuring-the-size-of-specimens-
viewed-with-a-microscope-and-adding-
scale-bars-to-photomicrographs January
24, 2024.
3. Linear Measurements (Micrometry).
Fig. 8. Making some changes in the fine
(n.d.). Nikon’s MicroscopyU.
adjustment knob to sharpen the focus.
https://www.microscopyu.com/microsco
py-basics/linear-measurements-
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4. To Calculate The Size Of An Organism
Under A Microscope –
modeladvisor.com. (n.d.).
Www.modeladvisor.com. Retrieved
February 4, 2024, from
Fig. 9. Viewing the specimen under Compound
https://www.modeladvisor.com/to-
microscope with a magnification of 400x.
calculate-the-size-of-an-organism-
under-a-microscope/.