Cell Biology

October 19, 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.That 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
identifying its cell structures.

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 was able to successfully discover, sketch and identify several
objects in different powers, such as cheek cells, elodea cell, and prepared slides of
amoeba and some bacterias.

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.

LITERATURE CITED

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