Cell Biology

October 18, 2017

Exploring Microscopy and Cell Anatomy and Diversity

Earlyn Joy V. Eniola

Science Department, College of Natural Sciences and Mathematics, Mindanao State University
General Santos City 9500

ABSTRACT
The microscope is designed to make objects visible that are too difficult or too small to
see with the unaided eye. This paper is all about the microscopes and cells. Microorganisms
and all other living organisms are classified as prokaryotes or eukaryotes. By performing wet
mount and staining of specimens and knowing the difference of eukaryotic and prokaryotic cell
by viewing under the microscope. Observing the cell structures, it differentiates that Eukaryotic
cells contain membrane bound organelles such as the nucleus, while prokaryotic cells do not.

INTRODUCTION
Understanding the nature if cell structure and function is important to an understanding
of organism. The cell is the fundamental biological unit according to cell theory, it is the
smallest and simplest biological structure possessing all the characteristics of the living
condition. All living condition. All living organism are composed of one or more cells, and
every activity taking place in a living organism is ultimately related to metabolic activities in
cells.
There are two general types of cells: prokaryotic and eukaryotic. Prokaryotic cells do
not have a membrane bound nucleus and their genetic material is loosely confined to a nuclear
area within the cell. Bacteria and archaea, including the cyanobacteria, are prokaryotes. All
other organisms are eukaryotes. The cells of eukaryotes have true, membrane-bound nuclei
containing their genetic material. Understanding the processes of life necessitates an
understanding of the structures and function of the cell.
Given the fundamental role played by cells in the organization if life, owe can readily
understand why the study of cells is essential to the study of life. Cells are very small and for
this reason, we cannot study them without using a microscope. The microscope has probably
contributed more than any other instrument to the development of biology as a science. There
are many kinds of microscope used today it differs primarily in the source and way light is
passed through the specimen to be viewed. The microscope is designed to make objects visible
that are too difficult or too small to see with the unaided eye.
In this laboratory, we observed selected specimens of eukaryotes and prokaryotes to
identify their cellular structures, and differentiate between prokaryotes and eukaryotes. We also
demonstrate using light and dissection microscopes to practice proper microscopy skills,
including making wet-mount slides and cell sizing. By observing, drawing, and classifying
bacteria, animal cells, plant cells, protozoans we learned about the cell structure. We also
learned about the differences and similarities of prokaryotic and eukaryotic cells.
MATERIALS AND METHODS
In this lab activity, the materials used are compound microscope, glass slides, cover
slips, lens paper , immersion oil for the Oil Immersion Objective viewing and the chemical
used for staining the cheek cell is methylene blue . And the specimens used to observe under
the microscope are the Bacillus subtilis smear, Shigella dysentenae smear, Cheek cells, Onion,
Elodea leaflet and a prepared slide of Amoeba.
A1. Parts of the microscope
The first thing we do is by getting the microscope in the bio stock room, making it sure
that we used our both hands in handling the microscope and placing it securely in the table with
arm facing towards us and we carefully identify each parts of the microscope and determining
its function.
A2. Determining the Magnifying Power
As we determined the magnification of each lenses by looking it in the engraved
magnification, we then calculate the total magnification of the specimen by multiplying the
magnification of the ocular lens and objectives lens.
A3. Image Orientation and Brightness
We prepare a cut letter e in the slide and place it in the stage in an upright position and
securing it by the used of the stage clips. Adjusting the letter e so that the light can easily
transmitted through the letter. We started the observation in the scanning lens while looking in
the eyepieces we carefully adjust the stage with used of the coarse adjustment knob until it
focuses. Observing it carefully what happens to the letter e by slowing moving the slide to the
left and towards us and determine the direction of the letter e appears. Then switch the lens
from scanning to lower objective observing the working distance , between the lens and the
slide and from the low power switch to high power objective observing the image and
brightness of the specimen.
A4. Size and the Diameter of the Field of View
As we determine the size and diameter of the field of view of each magnification, we
prepared first the slide with the millimeter gridlines and placed it onto the stage. Adjusting the
slide so that the light is transmitted through the gridlines. We started first with the scanning
lens, using the coarse adjustment knob to adjust the stage if it is upward or downward until the
gridlines are in focus, aligning the edge of the gridline with the stage of the field of view. And
we count the number of boxes that fits across the field of view and we recorded the value on
the worksheet. We also do the same in the Low power and high-power objective. As we
determined the size of the object we used this formula:
Size of the object= field diameter for that magnification
# of cells that fit across the diameter
A5. Depth of Field
In determining the depth of field we observed three colored threads if it is in focus in
LPO and if it still focuses in HPO.

B1. Bacteria
 We get a prepared bacterial slide in the bio stock room, the slides we obtained are
Bacillus subtilis smear and Shigella dysentenae smear. We observed it under OIO with
immersion oil and carefully observing its morphological structure and also identify its
components and what we observed we draw it in the worksheet.
B2. Animal Cells
To observed an animal cell, we prepared a wet mount of cheek cells, the first thing we
do is we get a glass slide, cover slip and a tooth pick, we used the toothpick to scrape the inside
of our cheek and we smear it in the empty glass slide and we drop a methylene blue stain into
it and allowing it in 2- 3 minutes. We first viewed the cells in the low power objective an switch
to high power objective, we draw the cheek cells and labelled the visible components.
B3. Plant Cells (Onion)
To observed a plant cell, we made a wet mount of an onion skin. Obtaining a thin piece
of an onion skin by taking an onion leaf and snapping it in two, carefully removing the thin
skin that separates from the convex side of the onion leaf. After that, placing the thin onion
skin on a clean slide and we add a few drops of iodine to stain the onion skin and allow the
stain to remain in few minutes and place a cover slip on the slide at the edge of the wet specimen
and carefully drop the cover slip to the specimen minimizing some air bubbles. and we start
viewing the specimen under LPO and HPO, under HPO what we draw what we observe and
labelled the visible components.
B4. Plant Cells (Elodea)
In obtaining a wet mount of an Elodea leaflet by placing the small leaflet on a clean
slide and cover with a cover slip after that we can view the Elodea cells under LPO and HPO
and observe and label the visible components. To enhance the visibility of the nucleus, use a
drop of iodine to stain.
B5. Eukaryotes- Protozoa, Algae, Slime molds
To observed a protozoa, algae and slime molds we get a prepared slide of a stained
amoeba and a paramecium in the bio stock room but the only available slide is amoeba only,
we draw the observed specimen and labelled the obvious parts.

RESULTS AND DISCUSSION

This part talks about the results and discussion of the conducted lab activity includes
the: Determining the Magnifying Power, Image Orientation and focusing, Diameter of the Field
of View, Depth of Field, Prokaryotic and Eukaryotic Cells.
Determining the Magnifying Power
In determining the magnification, the formula below was used. The magnification of
ocular lens and objective lens are also provided.
Formula: Total Magnification= Ocular Magnification X Objective magnification
Ocular Lens X Objective Lens = Total Magnification
The magnification of the ocular lens is 10X, the 4 objectives the Scanner, LPO, HPO,
OIO the magnification is 4X,10X, 40X, 100X. The total magnification of each objectives, the
scanning power is 40x, the Low Power is 100x, the High Power is 400x and the Oil Immersion
is 1000x.
Image Orientation and Focusing

Figure 1. Letter “e” on scanning power 40x

From the observation obtained, the final image of the specimen, which the position of “e” on the stage
is inverted. This is because due to the number of lenses within the microscope, or more specifically,
the number of focal point it has. Each focal point will invert the image once, meaning the microscope
with single lens will produce an inverted image. When the “e” slide id moved from left to right, the
image appear to move to the left since the image appears backwards when you look through the
ocular, and vice versa when moved towards you, the direction will be the same.

Figure 2. Letter “e” on Figure 3. Letter “e” on Figure 4. Letter “e” on

scanning power (40x) low power (100x) high power (400x)

As we conducted the laboratory, we observed that in scanner (40x) the letter e widely
observable in inverted position and in low power (100x) the letter e increase its size because it
magnification increases and when the letter “e” is observed using high power, the final image
become more blurred and the focus could be completely off due to the larger and blurred image.
We cannot see the whole part of “e”, yet we can only see only part of it. When this happen, the
limit of resolution of the microscope has been exceeded
In switching from low to high power the intensity of the light decreases as the
magnification increases. In adjusting the light intensity must be raised by turning the light
control knob clockwise because the light intensity decreases as the magnification increases.
The working distance decreases as the magnification increases.
 Determining the field of view is important for us to determine the size of our specimen.
When we observe the specimen, we must estimate its size by comparing it with the size of the
field.
Size of Field of view X Scanning power total magnification=Total magnification
Remember: The total magnification will depend on what objective’s diameter you want to get.
Scanning power diameter: 4.4 µm
Low power diameter: 1.76 µm
High Power diameter: 440 µm
Equation:
Estimated size of field of view: 4.4 mm
High power diameter:
Size of Field of view X Scanning power total magnification =Total magnification
= 4.4 mm X 40x
400x
After you have solved the diameter, convert it µm.
Note: 1mm = 1000 µm
0.44 mm x 1000 µm = 440 µm
It is important to know the diameter of the field of view, because we can use it to
determine or estimate the size of the organism you are viewing at that given magnification.
Depth of Field
We view three colored threads in low power and we tried if it is focused. And yes, it
was all focus. And it you view in on the microscope, from top to bottom, it is the arrangement:
Yellow, red, and blue. And if we are viewing organisms under LPO, we need to make sure that
it is focus before switching it to high power because microscopes are parfocal, once you have
focused carefully with the low power objective, the other objective will be nearly in perfect
focus.
Prokaryotic and Eukaryotic Cells

Figure 5. Bacillus subtilis Figure 6. Shigella dysentenae

smear (400x) smear (400x)

The figure 5 shows the Bacillus subtilis smear under HPO (400x). The Bacillus subtlilis
was a bacillus bacterial type because of its observable shape in rod the color of the cells are
violet and its arrangement was in pairs, individual and clusters.
The figure 6 shows the Shigella dysentenae smear under HPO (400x). The Shigella dysentenae
was a bacillus bacterial type because of its shape that is rod and the color of the cells are pinkish
and its arrangement was in pairs, individual and clusters.

Figure 7. Cheek Cell (400x)

The figure 7 shows the Cheek cell under HPO (400x). The observable structures in the cheek
cell are cell membrane, cytoplasm and nucleus.
 Figure 8. Onion Cell (400x)

The figure 8 shows the Onion Cell under HPO (400x). The visible components are cell wall
because it is a plant cell, cytoplasm and nucleus.

Figure 9. Elodea Cell (400x)

The figure 9 shows the Elodea Cell under HPO (400x). Elodea is an aquatic plant commonly
grown in freshwater aquaria. The cell structures may be difficult to see because of the three-
dimensional cell shape and the presence of a large central vacuole.

Figure 10. Amoeba (400x)

The figure 10 shows the Amoeba under HPO (400x). Amoeba has an irregular shape. Amoebas
do not seem to have a particular shape. We noticed a nucleus which is the control center of the
cell, aside from that we noticed the presence of the vacuole which is also large and circular in
shape located below the nucleus.
CONCLUSION AND PERSPECTIVE
The Microscope is a very powerful tool for understanding the structure and function of
tissues, and it is widely used in biomedical science courses, as well as in research and diagnostic
laboratories. In this lab activity, we observed not only the external features and functions of
the microscope, but also the specimens magnified through the microscope. We also made a
specimen through wet mount and drew observations carefully. I could successfully discover,
sketch and identify several objects in different powers, such as cheek cells, elodea cell, and
prepared slides of amoeba and some bacteria’s.
In conducting in this laboratory, we learned the similarities and differences between the
eukaryotic and prokaryotic cell. In eukaryotic cells there are plant and animal specimen to be
observed in plant cell is that there is a cell wall present while in the animal cell there is a cell
membrane. It is also observed that eukaryotic cells contain membrane bound organelles such
as the nucleus, while prokaryotic cells do not.

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