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 and its cell structure that is observed.

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 objectives diameter you want to get.

Scanning power diameter: 4.4 m

Low power diameter: 1.76 m

High Power diameter: 440 m

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 m.

Note: 1mm = 1000 m

0.44 mm x 1000 m = 440 m

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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http://dosequis.colorado.edu/Courses/MCDB3145/Docs/Karp-3-23.pdf

Axelrod D (2001) Total internal reflection fluorescence microscopy in cell biology. Traffic 2:764
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Thorn K. (2016) A quick guide to light microscopy in cell biology. Mol. Biol. Cell January 15,
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Murray (2004). Introduction to Botany 1st Edition. Pearson Education INC.

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