GROUP 6 - Laboratory Activity 1 Microscopy 1

POLYTECHNIC UNIVERSITY OF THE PHILIPPINES
COLLEGE OF SCIENCE
Department of Biology
BIOL 30035 GENERAL MICROBIOLOGY A.Y. 2021-2022
Name: Bagtas, Kiana Clarice L. Yr. & Sec: BS Biology 1-2

De Guzman, Rochele P. Professor: Dr. Lourdes Alvarez
Factor, Angeline P.
Luzuriaga, Ansley Mackenzie R.
LABORATORY LESSON
Activity No. 1
MICROSCOPY
I. Introduction
Microbiology has historically made the largest strides forward as new techniques are
produced and existing tools are improved. One of the tools that has improved over time is
the microscope. The microscope is the oldest and most basic tool used by microbiologists
to examine microorganisms (Madigan, 2018).
Some of nature's most fundamental processes take place on the microscopic scale, far
beyond the human ability to see with the naked eye, motivating the creation of technology
that allows people to look beyond these boundaries. People had discovered the basic
concept of an optical lens as early as the 4th century AD, and by the 13th century, they
were already using glass lenses to improve their eyesight and magnify objects like plants
and insects to better understand them. These simple magnifying glasses evolved into
advanced optical systems known as light microscopes, which allow people to see and
understand the microscopic world beyond the limits of their perception (Technology
Networks, 2022).
Microscopes are devices that magnify tiny objects in order to generate visual or
photographic pictures. A microscope must do three tasks: magnify an image, separate the
picture's features, and make these details visible to the naked eye or camera. Multiple-lens
microscope designs with objectives and condensers, as well as basic single-lens devices
that are typically handled, such as a magnifying glass, fall under this category.
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According to the Microbe Notes (2022), a compound light microscope has different
parts that have a different role in how it will work. It creates a picture by focusing light on the
specimen and magnifying it. Normally, the specimen is positioned near the microscopic
lens. The magnification of a microscope varies substantially depending on the type and
number of lenses that make up the instrument. A compound light microscope has a higher
magnification compared to a simple light microscope because it uses at least two sets of
lenses: an objective lens and an eyepiece.
Moreover, as stated by Pluta (1988), a compound light microscope does not just consist
of two lenses but is also commonly used to see plant and animal cell organelles, as well as
certain parasites like Paramecium, following staining with basic stains. Its functionality is
predicated on the ability to produce a high-resolution image, which is heavily dependent on
the microscope's optimal operation. This indicates that enough light will allow for effective
picture focusing, resulting in a high-quality image.
Furthermore, this instrument is solely based on the resolution of a lens, which is defined
as a lens's ability to distinguish tiny objects that are close together. The numerical aperture
of a light microscope's lens system and the wavelength of the light it uses define its
resolution; a numerical aperture is a description of the light wavelengths generated when
the specimen is lit (Prescot, 2018). Thus, the purpose of the compound light microscope is
to magnify pictures of specimens lit or producing light in the visible range of the spectrum,
as well as the neighboring ultraviolet and near-infrared parts of the spectrum.
Since the instrument "microscope" is used for precise observations and experiments, it
is a tool that necessitates correct handling and maintenance in order to produce reliable
results. Due to the delicate nature of a microscope, proper care and maintenance are
essential. There are many small items or details of objects that the unaided human eye
cannot see. It enlarges the image of such things, allowing them to be seen by the naked
eye. Microscopes are used to look at the forms of bacteria, fungus, parasites, and host cells
in various colors and unstained preparations (Lyons, 2021).
Thus, different types of microscopes, how to handle these delicate instruments, its parts
and functions are all part of microscopy. This enables researchers and scientists to perform
microscopic measurement and calibration using micrometry.
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Micrometry is the science of measurement of the dimensions of an object being

observed under the microscope. The method employs some special types of measuring
devices which are attached to or put into the microscope and observed.
Microscopy encompasses several sorts of microscopes, as well as how to manage this
sensitive equipment, as well as its parts and purposes. This allows researchers and
scientists to use micrometry to do microscopic measurements and calibration. Micrometry is
the study of determining the dimensions of an object seen under a microscope. The
procedure involves attaching or inserting special measuring devices into the microscope
and observing them (Sharda, 2020).
In science, microscopes have opened many doors. Microscopes enable scientists,
researchers, and students to uncover the presence of microbes, analyze cell structure, and
view the tiniest components of plants, animals, and fungus (Magnusopto, 2021). Without
the microscope, humanity would not have progressed as far as it has, and many diseases
would still be incurable. Microscopes are thus the foundation of biological research.
II. Objectives
Students are expected to learn more about microscopy as a result of the course
materials provided for this laboratory activity, as well as the assessments. In line with that,
this microscopy laboratory activity aims to: (1) identify different parts of the microscope and
their functions; (2) perform microscopic measurement and calibration using micrometry; (3)
determine the magnification for each of the following lenses by looking for the engraved
magnification; (4) calculate the total magnification of an object being viewed by multiplying
the magnification of the ocular lens and objective lens; (5) determine the value of the
calibration factor for the oil-immersion objective; (6) determine the size of the microbe cell
on the prepared slides under the oil-immersion objective; (7) determine the size of the other
microorganisms by observing the remaining prepared slides under oil immersion, and (8)
discuss the demonstration of the principles of proper use and care of a light microscope.
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III. Materials
Since this laboratory activity is done by groups and the students are in their own
homes due to the pandemic, materials such as microscopes, ocular micrometers, and other
laboratory equipment are not readily available, making it difficult for the students to use and
explore the instruments themselves. For these reasons, the researchers developed
alternative materials for this laboratory activity:
• Virtual microscope links
• Pencil
• Clean white paper
• Calculator
• Internet
If the students are on a laboratory set-up, the following materials should be available to
them in order to accomplish the laboratory activity for microscopy correctly:
• Compound microscope for magnification of cell sample
• Ocular micrometer to measure the size of the magnified object
• Stage micrometer to measure the smallest length of an object
• Cedar oil for immersion
• A cell sample to be observed
IV. Methods
The activity is intended for students to complete using mental visualization and thought
experiments. For students who do not have access to a laboratory, the internet, or other
relevant instructional materials, the module is sufficient. The students used the virtual
experiment links located in their professor's module for the first set of materials, in which
they compromised and identified things that would assist them conduct the laboratory
activity. Due to the online set-up, the researchers were unable to utilize an actual
microscope for this laboratory activity, so they used a virtual microscope instead.
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They went through the virtual experiment links one by one and discovered that two of
them were malfunctioning, while the other worked well. The students were able to learn
about several parts of the microscope by visiting that link. They virtually held the
microscope correctly in the base and arms before setting it on the lab bench with the
microscope's arm facing the researchers. The next step the students took was identifying
the microscope's components, which include the ocular lens, revolving nosepiece, objective
lenses, stage and stage clamp, iris diaphragm lever, condenser and adjustment knob,
coarse and fine adjustment knobs, slide movement knobs, and on/off knob/switch.
After learning about parts and functions of the ultimate tool used in microbiology, the
students then determined the magnifying power for each of the following lenses and looked
for the engraved magnification, then calculated the total magnification of the viewed object
by multiplying the magnification of the ocular lens and objective lens.
Despite the lack of an actual ocular micrometer and other laboratory tools for the final
part of the procedure, which involves calibrating the ocular micrometer and measuring
microorganisms, the researchers used the reference video given by their professor and
obtained some data from it.
Furthermore, the students moved on to the problem-solving section, which aided them
in the field of microbiological computation. It taught the students how to compute for known
distances, ocular division distances, the length of an organism being viewed, and many
other micrometry concepts. They distributed each problem, then used the calculator to
examine the problems and write solutions on a clean white piece of paper. Lastly, the
students entered their final solutions and answers into the word file.
V. Results
After the students have completed the procedures, it will be shown in this part what their
findings are after observing the various parts and functions of the microscope, how to
properly handle and care for the said instrument, as well as a thorough discussion to further
explain and familiarize the relationship between micrometry and microbiology.
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Figure 1. Learning Through Virtual Experiment
As per Figure 1, the researchers were able to list the basic components of a typical
microscope, describe the use of lens power and eyepiece powers, calculate the
magnification of a microscope based on the selected lens, and discuss the care and use of
a typical microscope after completing the exercise with the help of a virtual experiment link.
Moreover, by conducting practice and testing on the specified link, it demonstrated the
functions of each part after establishing the parts' identification: (a) ocular lens, a usually
10X lens which is at the upper end of the tube and the scopes that the students are using
are binocular with two eyepieces; (b) revolving nosepiece, the objective lenses are attached
below the nosepiece and this allows the user to change the magnification; (c) objective
lenses, the students found that the scopes either have 3 or 4 objective lenses attached to a
revolving nosepiece and the magnification is inscribed on each lens, and the power that are
available are: scanning (4X), low (10X) and high dry (43X). Some scopes (Swift) have the
oil immersion lens (100X); (d) stage and stage clamp, the slide rests on the stage and it is
held in place with the stage clamp while the moveable portion of the stage clamp should
only be touching one corner of the slide; (e) iris diaphragm lever, it is on the front edge
beneath the stage is a small lever that is used to adjust the contrast by regulating the
amount of light; (f) condenser and adjustment knob, the condenser condenses the light rays
into a stronger beam and the students used the adjustment knob located below and to the
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side of the stage to increase or decrease the light intensity; (g) coarse adjustment knob, on
each side of the scope was a large knob used to move the stage up and down to focus the
image and this knob is to be only used with the scanning and low power objective lenses;
(h) fine adjustment knob, it is located by the coarse adjustment knob and it allows for very
small changes to the height of the stage, as well as in increasing the sharp focus of an
image and the only knob to be used with high power; (i) slide movement knobs, these can
be found on each one side of the stage, there are two black or silver knobs that will be used
to move the slide, one knob will move the slide to the left and right, whereas the other knob
will move slide towards and away from the user. The scopes are parfocal, which means that
when the user have the object centered in the field of view on a lower power and then
change to a higher power, the image will remain in the center of the field of view; and (j)
On/Off Knob/Switch, this is located on side of the microscope or the top of the base in the
front end of the microscope.
Figure 2. Test Conducted Through Virtual Experiment
As shown in Figure 2, the students did not simply learn to store them in their short-term
memories, but also to retain them in their long-term memories as a series of tests were
given to challenge the students' knowledge and competence in the field of microscopy.
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Figure 3. Instruments Required for the Calibration

An ocular micrometer must be calibrated in order to measure the size of a microscopic
specimen. The instruments required for the procedure include a disused standard light
microscope with an ocular lens equipped with an ocular ruler, a slide micrometer, and a
slide with a specimen on it.
Figure 4. Calibration of the Ocular Micrometer

Since the image size of any specimen is magnified twice when viewed under a
microscope, researchers calibrated the ocular ruler first, and then did so for each objective
lens on the microscope. The objective lens magnified it first, followed by the ocular lens.
The objective lens was a 4x objective lens in this figure, whereas the ocular lens had a 10x
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magnification. As a result, the specimen was magnified four times, then ten times, for a total
of 40 times. That is the specimen's total magnification. The specimen was on a slide on the
microscope's stage, which is why the magnification is 40x. However, because the ocular
micrometer was positioned directly below the ocular lens, its image size was only enlarged
once by the ocular lens and was unaffected by the objective lens' magnification. As the
researchers did not alter the ocular lens, it remained unchanged in principle. As a result, the
calculated values for the ocular micrometer had to represent how the image size varies in
relation to the objective lenses, while the ocular ruler remained constant. After that, the
researchers modified it and recalculated it for each objective lens.
Figure 5. Calibration of the Stage Micrometer

The researchers first learned what a stage micrometer was and how to use one before
calculating the required values. This figure depicts the appearance of a common stage
micrometer. It is essentially a microscope slide with a little ruler carved into the surface.
This slide, like any other microscope slide, is mounted on the microscope stage. It consists
of many vertical lines when enlarged in the ruler's image. These vertical lines are also
known as division markers since the spaces between these lines are actually called
divisions, and the researchers focused on the gaps between the lines, indicating that the
divisions were significant. Additionally, these division markers, or lines, come in three
different sizes and lengths. The space or divisions between each of these divisions will be
the same regardless of their length, ensuring that they are evenly spaced apart. Therefore,
the researchers start by counting the division markers from left to right, beginning with zero,
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just like any other ruler, whether it is a US ruler or a metric ruler. After counting all of the
markers from left to right, regardless of size, the researchers discovered that there are 10
spaces or divisions between any two of the longest division markers, resulting in a total of
100 divisions. As a result, the researchers count these huge division marks in tens. By
including the mid-sized division markers, the researchers counted the number of divisions
by five. The stage micrometer has now been calibrated after some time. One division is
equal to 0.1 mm in length, which is quite a small amount, as stated in the upper right-hand
corner. There are 100 of these divisions, each of which is the same size; the distance
between any two division marks on this ruler is 0.01 mm. That is what the term "calibrated
ruler" refers to. Whether a person uses millimeters or micrometers, the result will be the
same as long as the person is consistent and utilizes the correct value. However, the
researchers used the term "one division" to refer to a length of 10 micrometers throughout
the procedure. The researchers simply multiplied the number of divisions, in this example,
100, by 10 μm because the micrometer used has 100 divisions and each division is said to
be 10 μm long. As a result, the division ruler measures 1000 micrometers in length, which
means that the division marker zero and the division marker 100 are separated by 1000
meters. One division equals ten meters, and the micrometer has 100 divisions, so its total
length is 1000 micrometers. The researchers used these two pieces of information to
assign a similar length unit to each division of the ocular micrometer, although this was
determined by the objective lens employed throughout the procedure.
Figure 6. Field of View on the Microscopic Slide

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The researchers used a slide micrometer to focus the image on the microscope slide,
then used a separate objective lens to focus the image. They began with the forex objective
lens and then mounted the stage micrometer on the stage microscope stage in this
scenario. They then used a 4x objective lens to focus it. They eventually noticed two rulers
in their line of vision. The stage micrometer was one, and the ocular micrometer was the
other. It is critical that the researchers keep track of which rulers belong to whom and where
the rulers originate. The researchers also observed that by using the stage X and Y
adjustment knobs, they could position both rulers to the left or right.
Figure 7. Calibration in a Total Magnification of 40x

After cleaning the slide, the researchers returned the values, which said that one stage
micrometer division equals 10 μm, and that there are 100 divisions, resulting in a total
length of 1000 μm for the stage micrometer. They have created two red dotted lines with
these two crucial values. Because the gap between these two red lines was 1000 μm and
covered the distance between 40 ocular divisions on the micrometer, the distance between
these two dotted lines was similar to 1000 micrometers and carried the red lines down to
the ocular micrometer. According to the researchers, 1000 micrometers is also equivalent
to 40 ocular divisions. They then calculated the length of one ocular division. Cross
multiplication was used to solve the problem. They assumed that 1000 μm is multiplied by
one ocular division. Then they divided the problem in half by 40 ocular divisions on both
sides. In order to arrive at the correct equation, the researchers eliminated necessary
values. The equation was changed to a thousand micrometers divided by 40. One arc
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division equals a thousand micrometers divided by 40 equals 25 micrometers, which gave
the researchers the number of micrometers. As a conclusion, the researchers used the 4x
objective lens to determine or calibrate the ocular micrometer. One artery division is equal
to 25 μm when using this objective lens.
Figure 8. Calculated Ocular Division at 40x Total Magnification

By determining the specimen size, the researchers substituted the stage micrometer with
a microscope slide containing a real specimen, and based on the ocular ruler, it is
estimated that it is around 6 divisions long. In addition, one division equals 25 μm, as per
the researchers' calculations. Because the specimen is 6 divisions long, the researchers
multiplied 6 divisions by 25 μm, resulting in a length of 150 μm.

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After that computation, the researchers reassembled the stage micrometer, swapped out
the 4x objective lens for a 10x objective lens, and repeated their calculations. The image on
the stage micrometer was significantly larger as a result of this, but the image on the ocular
micrometer remained unchanged. They compared the lengths of the two micrometers once
more. Remember that one division of the stage micrometer is 10 micrometers, and the
stage micrometer's length is 1000 micrometers. In this instance, the researchers compared
their lengths. A thousand micrometers were the length of the stage micrometer, and 100
micrometers equaled a thousand micrometers. The researchers divided the thousand
micrometers into a hundred ocular divisions in the ocular micrometer, which indicates that a
hundred ocular divisions equal a thousand micrometers. The researchers did the cross
multiplication again, so the equation would be 1 ocular division times a thousand
micrometers is equal to a hundred ocular divisions times whatever the number of
micrometers in one ocular division. Last time, they divided both sides by 40 to simplify, but
since 40 is not available this time, they divided both sides by 100 ocular divisions. Similarly,
the researchers performed some cancellation and simplified the equation, resulting in 1000
micrometers divided by 100, which equals 10 μm. So, with a 10x objective lens and a total
magnification of 100x, the answer in this example was 1 ocular division, which is equivalent
to 10 μm.

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The specimen is now around 14 divisions long after the researchers replaced the slide
micrometer with a specimen at 100x total magnification. It has a length of 12 to 26
micrometers. Therefore, 26 minus 12 divisions equals 14 divisions. Fourteen divisions
multiplied by ten micrometers equals 140 micrometers in length.

The researchers replaced the objective lens once more, removing the 10x and replacing
it with the 40x, as well as bringing back the stage micrometer. At 40x and 400x total
magnification, the stage micrometer image was now so big that the entire ruler does not fit
in this field of view, so the researchers did not rely on the fact that the stage micrometer
equals a thousand micrometers in length, rather they relied on the data wherein each stage
division is equal to 10 μm. They drew two dotted red lines again, this time deciding to place
them between ten stage divisions. They brought them all the way down to the ocular
micrometer. The distance between these two stages is 109 μm, which is equal to 10 stage
divisions multiplied by 10 micrometers. Moreover, the researchers calculated that 40 ocular
divisions are taken from 60 ocular divisions, then 60 is subtracted by 20, leaving them with
40 ocular divisions, based on this comparison. The 49 ocular divisions cover a distance of
100 micrometers. They conducted some cross multiplication again, using the same
equation as the two previous equations. One ocular division multiplied by 100 μm equals 49
ocular divisions multiplied by the number of micrometers in one ocular division. After that,
the researchers divided both sides of the equation by 40 ocular divisions to simplify it. The
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equation can be simplified to 100 m divided by 40, with 2.5 μm as the answer for one ocular
division.

The students have drawn some red dotted lines again and demonstrated that the red
dotted lines are between 45 and 20, indicating that the specimen is divided into 45–20
divisions. They concluded that each division is 25 micrometers long, making this red
specimen 62 ½ or 62.5 μm long in length. If the microscope has other objective lenses, for
example, many of them have a 100x objective lens, the operator must calibrate the ocular
micrometer with these other objective lenses.
Figure 13. Summary of Calculated Ocular Divisions

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After correctly calibrating the length of all ocular divisions using all of the different
objective lenses that came with the microscope, the students determined the number of
ocular divisions occupied by each cell sample and the average of the three measurements.
Unless the operator changes the lenses, the values associated with these appropriate
objective lenses will remain the same.
VI. Assessment: Critical Thinking and Learning Outcomes Review
Guide Questions:
1. What is the difference between magnification and resolution? Explain why they are
both important to be accounted in microscopy.
Magnification, often known as enlargement, is the ability to amplify small objects in
order to make them appear larger, such as making a microscopic organism visible. In
addition, magnification is the function of a two-lens system: (1) the ocular lens that can be
found in the eyepiece and (2) the objective lens that can be found in a revolving nosepiece.
Resolution is the shortest distance among points that may nonetheless be visually
distinguished as discrete. Magnification and resolution are each vital due to the fact that the
purpose of a microscope is to enlarge functions to the point where new details may be
resolved. Thus, magnification refers to the ability to magnify small items, such as a
microscopic organism, whereas resolution refers to the ability to differentiate two objects
from one another. In reality, microbiologists rely on resolution more because they want to
be able to distinguish between microbes or sections of microbes (Biology LibreTexts, 2021).
A viewer must first zoom to a degree where resolution becomes relevant before being able
to differentiate between two objects under a microscope.
2. Aside from other control and adjustment knobs, why it is equally important to
adjust the iris diaphragm and condenser while observing specimens under the
microscope?
Since the iris diaphragm is used to alter the contrast by regulating the amount of
light, and the condenser condenses the light rays into a stronger beam, it is critical to adjust
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the iris diaphragm and condenser equally while studying specimens under a microscope.
Furthermore, in the pursuit of a perfect image, proper use of the condenser, which on most
microscopes contains an iris diaphragm, is important. When the condenser is raised to a
point slightly below the stage, a spotlight effect is created on the specimen, which is
important when using higher magnification lenses with small apertures (Ahern, n.d.).
3. How the total magnifying power of the lenses in the microscope is determined?
Give some examples.
Light from an object travels through a biconvex lens and is bent (refracted) towards
the individual's eye in simple magnification. It gives the impression that it came from a
much larger object. As stated by Science Learning Hub (2020), modern microscopes, rather
than having only one lens, have a variety of them. They have an objective lens (which sits
close to the object) and an eyepiece or ocular lens (which sits closer to the eye). Both of
these factors add to the object's magnification. The eyepiece lens typically magnifies 10x,
whereas the objective lens usually magnifies 40x. Microscopes normally come with a pair of
objective lenses that can be swapped out to modify the magnification. For instance,
by multiplying the magnifying powers of the objective lens and the ocular (40x times 10x),
the overall magnifying power of the microscope would be a total magnification of 400x.
Adding extra lenses to a microscope does not change the core principle of how it magnifies,
but it does allow for higher magnifications and a higher-quality image.
4. Why are the lengths of the cells usually approximated when we record their values
for length?
The length of the cells is usually approximated when we record their values for length
because these organisms are not round. The microscope may be used to measure the size
of very small things as well as examine them. The metric system, which is a decimal
system based on the meter, is used to convey microscopic measurements. Thus, the length
of an organism is usually approximated in a computation, considering organisms undergo
distinct adaptations and come in varying sizes and forms depending on their environment
and unique traits based on their genus or kingdom. Since the cells are growing, these
length variations arise (increasing in length). Some cells have recently divided, resulting in
the formation of shorter cells.
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5. Do the calibration factor computed varies according to different magnification

(objective lenses being used)? Explain.
Yes, the calibration factor computed varies according to different magnifications when
objective lenses are used because there are different powers that can be used under the
objective lenses. Each magnification has a distinct conversion factor. As a result, while
utilizing a reticule for the first time, the scale must be calibrated by focusing on a second
micrometer scale (a stage micrometer) placed immediately on the stage.
Problem-Solving:
PROBLEM #1
a) A commercial stage micrometer has a total length of 1000 µm and is divide into 100
equal divisions. What is the length of each stage division in millimeters? Show your
solution.
GIVEN:
• Total length of commercial stage micrometer = 1000 µm

• Stage divisions = 100 equal divisions
ASKED:
Length of each stage division in millimeters (mm) = ?
FORMULA:
𝑇𝑜𝑡𝑎𝑙 𝑙𝑒𝑛𝑔𝑡ℎ 𝑜𝑓 𝑐𝑜𝑚𝑚𝑒𝑟𝑐𝑖𝑎𝑙 𝑠𝑡𝑎𝑔𝑒 𝑚𝑖𝑐𝑟𝑜𝑚𝑒𝑡𝑒𝑟

𝑆𝑡𝑎𝑔𝑒 𝑑𝑖𝑣𝑖𝑠𝑖𝑜𝑛𝑠
= 𝐿𝑒𝑛𝑔𝑡ℎ 𝑜𝑓 𝑒𝑎𝑐ℎ 𝑠𝑡𝑎𝑔𝑒 𝑑𝑖𝑣𝑖𝑠𝑖𝑜𝑛 𝑖𝑛 𝑚𝑖𝑙𝑙𝑖𝑚𝑒𝑡𝑒𝑟𝑠
SOLUTION:
1000 µm
= 10 µm
100
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Convert 10 µm to mm:
1 𝑚𝑚
10 µm × = 𝟎. 𝟎𝟏 𝒎𝒎
1000 µm
ANSWER:
A commercial stage micrometer has a total length of 1000 µm and 100 equal
divisions, each of which is 0.01 mm long.
b) Twelve (14) ocular divisions coincide with three stage divisions. Assuming that the
graduations of the stage micrometer are spaced 10 µm, what is the known distance
between the three stage divisions in millimeters? Show your solution.
GIVEN:
• Ocular divisions = 12
• Stage divisions = 3
• Graduations of the stage micrometer = 10 µm
ASKED:
Known distance between the three stage divisions in millimeters (mm) = ?
FORMULA:
Stage divisions × Graduations of the stage micrometer = Known distance
SOLUTION:
3 × 10 µm = 30 µm
Convert 30 µm to mm:
1 𝑚𝑚
30 µm × = 𝟎. 𝟎𝟑 𝒎𝒎
1000 µm
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ANSWER:
Assuming the stage micrometer's graduations are spaced 10 µm apart, the known
distance between the three stage divisions in millimeters is 0.03 mm.
c) With the same given information on b), what would be the distance of one ocular
division? Show your solution.
GIVEN:
• Ocular divisions = 12
• Stage divisions = 3
• Graduations of the stage micrometer = 10 µm
• Known distance between three stage divisions in millimeters = 0.03 mm
ASKED:
Distance of one ocular division = ?
FORMULA:
𝐾𝑛𝑜𝑤𝑛 𝑑𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑏𝑒𝑡𝑤𝑒𝑒𝑛 𝑡ℎ𝑟𝑒𝑒 𝑠𝑡𝑎𝑔𝑒 𝑑𝑖𝑣𝑖𝑠𝑖𝑜𝑛𝑠 𝑖𝑛 𝑚𝑚

𝐷𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑜𝑓 𝑜𝑛𝑒 𝑜𝑐𝑢𝑙𝑎𝑟 𝑑𝑖𝑣𝑖𝑠𝑖𝑜𝑛 =
𝑁𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑑𝑖𝑣𝑖𝑠𝑖𝑜𝑛𝑠 𝑜𝑛 𝑜𝑐𝑢𝑙𝑎𝑟 𝑚𝑖𝑐𝑟𝑜𝑚𝑒𝑡𝑒𝑟
SOLUTION:
0.03 𝑚𝑚
= 0.0025 𝑚𝑚
12
Convert 0.0025 mm to µm:
1000 µm
0.0025 𝑚𝑚 × = 𝟐. 𝟓 µ𝐦
1 𝑚𝑚
ANSWER:
With the same given information from letter b, the distance of one ocular division is
2.5 µm.
COLLEGE OF SCIENCE
PROBLEM #2
d) Based on the image, how many ocular divisions and stage divisions respectively it
takes for them to exactly and accurately coincide with each other?
Based on the given illustration, it would take 4.2 ocular divisions to coincide with 2
stage micrometers accurately and unquestionably. The physical length of the specimen
that will be examined under the microscope may be calculated using the given divisions.
e) Based on a certain magnification the image was observed, if one stage division
corresponds to 0.1 mm distance, what would be distance of each ocular division?
If one stage division corresponds to a 0.1 mm distance, the measured distance of

one ocular micrometer is about 47.62 µm. The measured distance was evaluated by
multiplying the quantity of stage micrometer division by its distance and dividing it by the
number of ocular micrometer divisions.
GIVEN:
• 100 µm × 2 = 2 stage micrometer divisions

• 2 stage micrometer divisions = 200 µm
• 4.2 ocular micrometer division = 200 µm
ASKED:
Distance of each ocular division = ?

COLLEGE OF SCIENCE
FORMULA:

𝐷𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑜𝑓 𝑜𝑛𝑒 𝑜𝑐𝑢𝑙𝑎𝑟 𝑑𝑖𝑣𝑖𝑠𝑖𝑜𝑛 =
𝑁𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑑𝑖𝑣𝑖𝑠𝑖𝑜𝑛𝑠 𝑜𝑛 𝑜𝑐𝑢𝑙𝑎𝑟 𝑚𝑖𝑐𝑟𝑜𝑚𝑒𝑡𝑒𝑟
SOLUTION:
2 stage divisions = 0.1 mm × 2
2 stage divisions = 0.2 mm
0.2 𝑚𝑚
One division on ocular micrometer in mm =
4.2
= 0.04762 mm
Convert mm to µm:
1 Ocular division (µm) = 47.62 µm
ANSWER:
If one stage division corresponds to a 0.1 mm distance, the distance of each ocular
division would be 47.62 µm.
PROBLEM #3
f) What is the size of the nucleus in micrometers?
The nucleus size is determined by the size of the cell in which it is enclosed, with a
nucleus generally comprising roughly 8% of the total cell volume. The size of the nucleus in
the illustration was based on the given reference in the module. It is converted into
COLLEGE OF SCIENCE
micrometers, which is the measurement used in studying microorganisms by multiplying it
to take measurements of 0.1 mm to 1000 µm (which equates to 1 mm). Therefore, the
answer to the solution of the size of the nucleus is 100 µm.
GIVEN:
1
• Size of the nucleus = 10 𝑚𝑚 or 0.1 mm
• 1 mm = 1000 µm
ASKED:
Size of the nucleus in micrometers (µm) = ?
FORMULA:
Size of the nucleus in µm = Size of the nucleus in mm × 1000 µm
SOLUTION:
0.1 mm × 1000 µm = 100 µm or 100 micrometers
ANSWER:
The size of the nucleus in micrometers is 100 µm.
g) What is the size of the cell in micrometers?
Cells can vary from 1 micrometer (µm) to hundreds of micrometers in diameter.

Within a cell, a DNA double helix is approximately 10 nanometers (nm) wide, whereas the
cellular organelle called the nucleus that encloses this DNA can be approximately 1000
times bigger (about 10 µm). Using the given reference, the size of the cell in micrometers
will be computed. The known value of the cell in millimeters (mm) is 0.5, which will be
multiplied by 1000 µm (which equates to 1 mm). Therefore, the answer to the solution of
the size of the cell in micrometers is 500 µm.
GIVEN:
1
• Size of the cell = 2 𝑚𝑚 or 0.5 mm
• 1 mm = 1000 µm
COLLEGE OF SCIENCE
ASKED:
Size of the cell in micrometers (µm) = ?
FORMULA:
Size of the cell in micrometers (µm) = Size of the cell in mm × 1000 µm
SOLUTION:
0.5 mm × 1000 µm = 500 µm
ANSWER:
The size of the cell in micrometers is 500 µm.
PROBLEM #4
Under microscopic observation of a string of rod-shaped bacteria, the researchers

will observe how many individual cells the string is comprised of. Bacillus (genus Bacillus),
or bacilliform bacterium, is a rod-shaped, gram-positive, aerobic or anaerobic
microorganism found in soil and water. All cylindrical or rod-like bacteria are referred to as
bacillus.
h) Based on your microscopic observation, how many individual cells thus the string is
comprised of?
COLLEGE OF SCIENCE
Based on the photomicrograph, the string of a rod-shaped structure consists of

seven (7) individual cells. The number of cells was calculated by getting the approximate
size of a single cell and estimating the number of cells that could fit into the given example.
i) What is approximate combine cell length of the cells?
Based on the given scale, a single cell is around 5 µm. Using the standard size of a
single cell, its size, which is 5 µm will be multiplied by 7 (the computed number of individual
cells) and the answer will be 35 µm. Therefore, the combined length of the cells in the
illustration is 35 µm.
Remember: Always measure the cell length base on the widest/longest midsection of the
cell!
PROBLEM #5
Given:
• The used ocular objective while taking this image, has magnifying power of 6x
• The used objective while taking this image, has a magnifying power of 100x
• The used stage micrometer has spaces graduation of 0.01 mm each
• Ten (10) graduations on the ocular micrometer coincided with two (2) graduations
on the stage micrometer
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j) What is the total magnification of the image under microscope? Show your solution.
GIVEN:
• Magnification of the ocular lens = 6x

• Magnification of the objective lens = 100x
• Each space graduation of the stage micrometer = 0.01 mm
• Ocular micrometer = 10 graduations
• Stage micrometer = 2 graduations
ASKED:
Total magnification of the image under microscope = ?
FORMULA:
Magnification of the ocular lens × Magnification of the objective lens = Total magnification of
the image under microscope
SOLUTION:
6x × 100x = 600x
ANSWER:
Therefore, the total magnification of the image under the microscope is 600x.
k) What is the distance of one ocular division? Show your solution.
GIVEN:

• Total magnification of the image under the microscope = 600x
COLLEGE OF SCIENCE
ASKED:
Distance of one ocular division = ?
FORMULA:
• Graduations of stage micrometer × Each space graduation of the stage micrometer

= Known distance
• 𝐷𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑜𝑓 𝑜𝑛𝑒 𝑜𝑐𝑢𝑙𝑎𝑟 𝑑𝑖𝑣𝑖𝑠𝑖𝑜𝑛 = 𝑁𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑑𝑖𝑣𝑖𝑠𝑖𝑜𝑛𝑠 𝑜𝑛 𝑜𝑐𝑢𝑙𝑎𝑟 𝑚𝑖𝑐𝑟𝑜𝑚𝑒𝑡𝑒𝑟
SOLUTION:
Solve first for the known distance:
2 × 0.01 mm = known distance of 0.02 mm
Then, solve now for the distance of one ocular division:
0.02 𝑚𝑚
= 2 × 10−3 𝑜𝑟 0.002 𝑚𝑚
10
Convert 0.002 mm to µm:
1000 µm
0.002 𝑚𝑚 × = 𝟐 µ𝐦
1 𝑚𝑚
ANSWER:
The distance of one ocular division is 2 µm.
l) What is approximate cell length of Cell A?
GIVEN:

COLLEGE OF SCIENCE
• Distance of one ocular division = 2 µm
ASKED:
Approximate cell length of Cell A = ?
FORMULA:
• Length of organism = number of ocular divisions occupied × calibration factor for

one ocular division
SOLUTION:
1.8 × 2 µm = 3.6 µm
ANSWER:
For Cell A, its approximate cell length is 3.6 µm.
m) What is approximate cell length of Cell B?
GIVEN:

• Approximate cell length of Cell A = 3.6 µm
ASKED:
Approximate cell length of Cell B = ?

COLLEGE OF SCIENCE
FORMULA:

one ocular division
SOLUTION:
1.3 × 2 µm = 2.6 µm
ANSWER:
For Cell B, its approximate cell length is 2.6 µm.
n) What is approximate cell length of Cell C?
GIVEN:

• Approximate cell length of Cell B = 2.6 µm
ASKED:
Approximate cell length of Cell C = ?
FORMULA:

one ocular division
COLLEGE OF SCIENCE
SOLUTION:
1.6 × 2 µm = 3.2 µm
ANSWER:
For Cell C, its approximate cell length is 3.2 µm.
o) What is approximate combine cell length of Cell A and Cell B? Show your solution.
GIVEN:

• Approximate cell length of Cell B = 2.6 µm
• Approximate cell length of Cell C = 3.2 µm
ASKED:
Approximate cell length of Cell A and Cell B = ?
FORMULA:
• Cell A + Cell B
SOLUTION:
3.6 µm + 2.6 µm = 6.2 µm

COLLEGE OF SCIENCE
ANSWER:
When the approximate cell lengths of Cell A and Cell B are combined, it results in
6.2 µm.
Remember: Always measure the cell length on the longest middle section of the cell!
VII. Conclusion
The students were able to complete all of the objectives specified in the report at the
completion of this laboratory activity. Through the virtual experiment link, they first studied
and explored the parts of the microscope and its functions. The students were able to learn
not only about the microscope's parts and functions, but also how to properly use and
maintain the microscope in order to obtain accurate findings. They also used micrometry to
undertake microscopic measurement and calibration throughout the instructional material's
procedures. When operating and using a compound microscope, each component
complements the others. Therefore, some components will not yield accurate findings if
others are malfunctioning. Before utilizing the microscope, it is critical to ensure that it is in
good condition since it would be ineffective otherwise. Furthermore, the students computed
the total magnification of the object being observed by multiplying the magnification of the
ocular lens and objective lens, which they determined by estimating the magnifying power
of each. Lastly, there are three objective lenses for calibrating measurements with an ocular
micrometer and measuring microorganisms: the first has a 4x objective lens, the second
has a 10x objective lens, and the third has a 40x objective lens. When all of those are
multiplied by a 10x ocular lens magnification, the total magnification of each is 40x, 100x,
and 400x, respectively. After calibrating all the ocular divisions with the various objective
lenses that came with the microscope, the students discovered that it has a length of 25
micrometers at 40x total magnification. At 100x, it resulted in 10 micrometers and, lastly, at
a total magnification of 400x, the length is 2.5 micrometers. Microscopy is therefore crucial
to study for all of us since it allows us to observe the tiniest aspects of the world, such as
bacteria, small structures within larger objects, and even the molecules that make up all
matter. In many ways, the ability to see things that are ordinarily unseen enriches people's
lives.
COLLEGE OF SCIENCE
VIII. References
Ahern, H. (n.d.). The Microscopic World | Microbiology: A Laboratory Experience. Lumen

Learning. https://courses.lumenlearning.com/suny-microbio-labexperience/chapter/the-
microscopic-world/
Biology LibreTexts. (2021, January 4). 3.1D: Magnification and Resolution.
https://bio.libretexts.org/Bookshelves/Microbiology/Book%3A_Microbiology_(Boundless
)/3%3A_Microscopy/3.1%3A_Looking_at_Microbes/3.1D%3A_Magnification_and_Res
olution
Bolduc, G. (2017, February 9). Calibration of Microscopic Ocular Micrometer [Video].
YouTube. https://www.youtube.com/watch?v=HaqgCtA-ioI
Microbe Notes. (2022). Light Microscope - Definition, Principle, Types, Parts, Labeled
Diagram, Magnification. https://microbenotes.com/light-microscope/
Pluta, M. (n.d.). Advanced Light Microscopy. https://www.ohsu.edu/advanced-light-
microscopy-core
Prescot, L. (2018). Microbiology.
https://science.umd.edu/CBMG/faculty/wolniak/wolniakmicro.html
Science Learning Hub. (2020, June 2). How microscopes magnify.
https://www.sciencelearn.org.nz/resources/496-how-microscopes-magnify
Technology Networks. (2022). Analysis and Separations.
https://www.technologynetworks.com/analysis/articles/an-introduction-to-the-light-
microscope-light-microscopy-techniques-and-applications-351924

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POLYTECHNIC UNIVERSITY OF THE PHILIPPINES

BIOL 30035 GENERAL MICROBIOLOGY A.Y. 2021-2022

Name: Bagtas, Kiana Clarice L. Yr. & Sec: BS Biology 1-2

Micrometry is the science of measurement of the dimensions of an object being

Figure 1. Learning Through Virtual Experiment

Figure 2. Test Conducted Through Virtual Experiment

Figure 3. Instruments Required for the Calibration

Figure 4. Calibration of the Ocular Micrometer

Figure 5. Calibration of the Stage Micrometer

Figure 6. Field of View on the Microscopic Slide

Figure 7. Calibration in a Total Magnification of 40x

Figure 8. Calculated Ocular Division at 40x Total Magnification

Figure 9. Calibration in a Total Magnification of 100x

Figure 10. Calculated Ocular Division at 100x Total Magnification

Figure 11. Calibration in a Total Magnification of 400x

Figure 12. Calculated Ocular Division at 400x Total Magnification

Figure 13. Summary of Calculated Ocular Divisions

VI. Assessment: Critical Thinking and Learning Outcomes Review

5. Do the calibration factor computed varies according to different magnification

• Total length of commercial stage micrometer = 1000 µm

Length of each stage division in millimeters (mm) = ?

𝑇𝑜𝑡𝑎𝑙 𝑙𝑒𝑛𝑔𝑡ℎ 𝑜𝑓 𝑐𝑜𝑚𝑚𝑒𝑟𝑐𝑖𝑎𝑙 𝑠𝑡𝑎𝑔𝑒 𝑚𝑖𝑐𝑟𝑜𝑚𝑒𝑡𝑒𝑟

Known distance between the three stage divisions in millimeters (mm) = ?

Stage divisions × Graduations of the stage micrometer = Known distance

Distance of one ocular division = ?

𝐾𝑛𝑜𝑤𝑛 𝑑𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑏𝑒𝑡𝑤𝑒𝑒𝑛 𝑡ℎ𝑟𝑒𝑒 𝑠𝑡𝑎𝑔𝑒 𝑑𝑖𝑣𝑖𝑠𝑖𝑜𝑛𝑠 𝑖𝑛 𝑚𝑚

Convert 0.0025 mm to µm:

If one stage division corresponds to a 0.1 mm distance, the measured distance of

• 100 µm × 2 = 2 stage micrometer divisions

Distance of each ocular division = ?

𝐾𝑛𝑜𝑤𝑛 𝑑𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑏𝑒𝑡𝑤𝑒𝑒𝑛 𝑡ℎ𝑟𝑒𝑒 𝑠𝑡𝑎𝑔𝑒 𝑑𝑖𝑣𝑖𝑠𝑖𝑜𝑛𝑠 𝑖𝑛 𝑚𝑚

2 stage divisions = 0.1 mm × 2

2 stage divisions = 0.2 mm

1 Ocular division (µm) = 47.62 µm

f) What is the size of the nucleus in micrometers?

Size of the nucleus in micrometers (µm) = ?

Size of the nucleus in µm = Size of the nucleus in mm × 1000 µm

0.1 mm × 1000 µm = 100 µm or 100 micrometers

The size of the nucleus in micrometers is 100 µm.

g) What is the size of the cell in micrometers?

Cells can vary from 1 micrometer (µm) to hundreds of micrometers in diameter.

Size of the cell in micrometers (µm) = ?

Size of the cell in micrometers (µm) = Size of the cell in mm × 1000 µm

0.5 mm × 1000 µm = 500 µm

The size of the cell in micrometers is 500 µm.

Under microscopic observation of a string of rod-shaped bacteria, the researchers

Based on the photomicrograph, the string of a rod-shaped structure consists of

i) What is approximate combine cell length of the cells?

• Magnification of the ocular lens = 6x

Total magnification of the image under microscope = ?

k) What is the distance of one ocular division? Show your solution.

• Magnification of the ocular lens = 6x

Distance of one ocular division = ?

• Graduations of stage micrometer × Each space graduation of the stage micrometer

Solve first for the known distance:

2 × 0.01 mm = known distance of 0.02 mm

Then, solve now for the distance of one ocular division:

Convert 0.002 mm to µm:

The distance of one ocular division is 2 µm.