Name: Bayani, Koreen C.

Date performed: January 28, 2019 Score:

Section: Botany 111.1 – BC Date submitted: January 31, 2019

EXERCISE 1
Wet Mount Preparation

Introduction
In everyday life, the particles that surround people are mostly naked to the human eyes. To
be able to view these tiny particles is with the use of a microscope. The compound microscope
requires skills necessary for the understanding of the makeup of organisms and how they react in
certain environment. In order to accurately look at these organisms under a microscope, students
need to learn how to properly prepare a wet mount. A wet mount is made by placing a fluid solution
on a slide, suspending a specimen in a solution, and then covering the specimen and the solution
with a cover slide, all done in careful execution. This method is widely used in areas of biology,
botany, and zoology as it increases the specimens’ translucency.

Objectives
The main objectives of the exercise performed by the students were to learn how to
properly make a wet mount slide to see through the microscope and know what to observe and
describe. Furthermore, it aimed to help students accurately sketch, and identify the different
organelles and other cellular components of the cells examined as well as describe the similarities
and differences in terms of the structure of the plant cells.

Procedures:
Wet Mount of Water Plant (Hydrilla)
In preparing the wet mount for the water plant, a drop of water was added on a dry, clean
glass slide. Using the needle, the tip of water plant leaf which had been cut off was carefully placed
in the water, followed by a clean cover slip positioned on top. The slide was viewed on the
microscope using the scanner with 40X magnification to easily locate and focus on the specimen
as it offers the widest field of visible range. Once the specimen was located, the lower power
objective was used to observe the cell. Adjusting the coarse fine knob, the high power objective
was also utilized to view the specimen and observe the cytoplasmic streaming in a more close-up
look which provided clearer details. The few water plant cells were drawn at different
magnifications labeling the observed parts such as the nucleus, cytoplasm, cell wall, and the
chloroplasts.
Wet Mount of Potato

The following procedure consisted of two parts: the unstained specimen and the stained
specimen. In preparing the slide for the potato, a drop of water was placed on a microscope slide.
A slice of very thin piece of potato was put on the drop of water. The coverslip was placed on the
potato. As the coverslip wobbled, a piece of thin potato layered was sliced again because it was
too thick the first time. The specimen was observed under the microscope. The plant cells were
located using 40X magnification, then changed to 100X magnification for a more detailed image,
then turned to 400X magnification. The specimen was observed carefully to be able to draw a few
potato cells and correctly identify and label the nucleus, cytoplasm, cell wall, and amyloplasts.
The second part of the procedure was the replication of the first, with the addition of iodine
solution before the cover slip was placed on top. The reason for adding iodine was to stains starch
inside amyloplasts (look like bubbles).

Wet Mount of Onion Bulb

In a wet mount slide for the unstained onion bulb, a dry, clean slide was obtained and a
drop of water was added at the center of the microscopic slide. A very thin layer of onion was
carefully laid in the water. The drop of water was used to flatten the membrane. Once the specimen
was placed, the microscopic cover slip was set on top of the membrane and pressed gently using a
needle to remove air bubbles. A blotting paper was positioned on one side of the slide to drain
excess water solution. The slide was observed on the microscope using the scanner with 40X
magnification to locate the specimen. For a clearer image of the specimen, the objective was
adjusted to the lower power objective with 100X magnification. In order to zoom in and focus
more, the objective was changed into the higher power objective.

The second part of the procedure was also a replication of the first, with the addition of
iodine solution before the cover slip was placed on top. The differences between the stained and
unstained onion bulb cells were noted.

Results and Discussion

The lab exercise presents an introduction to the compound microscope followed by the
proper preparation of a wet mound slide of different specimens. Learning how to prepare a wet
mount for the microscope was essential for all procedures. The first procedure consisted of
preparing a wet mount of hydrilla, a known water plant. The specimen was observed in three
objectives with an increased magnification: scanner, low power objective, high power objective.
Figure 1. Water plant cell, Figure 2. Water plant cell, Figure 3. Cytoplasmic
40X magnification 100X magnification streaming in water plant
cell, 400X magnification

The images above show the water plant as viewed in the microscope under different
magnifications. The results concluded that the objective with the lowest magnification is used
when getting a specimen in focus as it is not too zoomed in so aligning the side and the lens is
easier. Also, by using the lowest magnification, the specimen is easier to locate, center, and focus
in on. Once the specimen is in sight, a gradual increase in the magnification reaching the high
power objective produces a very fine detail of the specimen and the cell shape, cell orientation,
parts and movements can be observed.

As a eukaryotic cell, the hereditary material of the cell or the DNA of the plant cell is
enclosed within the nucleus. It is also considered as the control center of the cell. The cytoplasm
is the gel – like matrix inside the cell membrane which constitutes all other organelles.
Furthermore, the most important distinctive structure of the plant cell is the presence of a cell wall.
It forms the outer lining of the cell thus providing support and rigidity. The chloroplasts aid in
carrying out the process of photosynthesis to produce food. These organelles carry out specific
functions necessary for the survival and normal operations of the cell.

A movement within the cell was also observed under the high power objective. In figure
3, the image of the cytoplasmic streaming is seen. Cytoplasmic streaming, also called as
protoplasmic streaming, is a movement of cytoplasm in various organisms including plants
(Ashley & Williamson, 1982). It assists in the delivery of nutrients, metabolites, organelles and
other materials to all parts of the cell and plays key roles in biological processes such as movement
of chloroplasts in plants (Kadota, et. al., 2009).

In the second procedure, a slice of very thin piece of potato was obtained, placed on a wet
mount, and covered by a cover slip. The specimen was viewed under different magnification
starting from 40X magnification, then 100X magnification, and followed by 400X magnification.
Figure 4. Potato cell, 40X Figure 5. Potato cell, Figure 6. Potato cell,
magnification 100X magnification 400X magnification

The images above show the unstained specimen in different objectives. The results
concluded that cells observed under the microscope lack color and contrast. This made it difficult,
if not possible, to detect important cellular structures and their distinguishing characteristics.

Figure 7. Potato cell, 40X Figure 8. Potato cell, Figure 9. Potato cell,
magnification 100X magnification 400X magnification

The procedure was recreated for the second time with the addition of iodine solution on
the wet mount before placing the cover slip. The results showed that using a stained specimen was
easier to locate rather than the unstained. Furthermore, the stain highlighted the outlines of the cell
and provides a stronger contrast and stains the starch inside the amyloplasts which made it easier
to differentiate and have a better look. Thus, a better visualization of cells and cells parts under
that microscope were observed.

By using iodine solution, the nucleus, cytoplasm, cell wall, and amyloplasts are easier to
view and identify. As mentioned above, nucleus contains the cell’s hereditary information and
cytoplasm gel-like substance within the cell membrane that contains water, enzymes, organelles,
and other various organic molecules. The cell is the rigid outer covering of the cell which protects
and gives it shape. In addition to having these, amyloplasts were also found on the potato cells.
These are organelles where starch is made and stored. Their functions are degradation and
synthesis of starch and ore it in the stroma (Lppez, 2016).

The third procedure consisted of two parts: the unstained and stained layer of onion bulb.
The unstained slide was observed on the microscope using the scanner with 40X magnification to
locate the specimen. For a clearer image of the specimen, the objective was adjusted to the lower
power objective with 100X magnification. In order to zoom in and focus more, the objective was
changed into the higher power objective.

Figure 10. Onion bulb Figure 11. Onion bulb Figure 12. Onion bulb
cell, 40X magnification cell, 100X magnification cell, 400X magnification

The images above show the unstained specimen in different objectives. The result
concluded that cells observed under the microscope were easier to visualize than the unstained
slice of potato. The onion bulb possesses a natural dye which gives it more contrast than the potato.
Some parts are readily seen such as the cell wall, nucleus and cytoplasm.

Figure 13. Onion bulb Figure 14. Onion bulb Figure 15. Onion bulb
cell, 40X magnification cell, 100X magnification cell, 400X magnification
 However, they still lack color thus an iodine solution was added to improve the contrast of
the faint images. In the images above, the parts are clearly seen even under 100X magnification
and an even detailed view under the 400x magnification. The spots present in the images are the
nucleus which contains the DNA. The highlighted outlines that surround the nucleus is the cell
wall which protects and gives shape to the cell. Meanwhile, the space inside or the gel-like
substance is called cytoplasm.

The results, in general, concluded that some parts of the plant cell present in the water plant
was not observes in the other two specimen such chloroplast. Aside from that, the amyloplasts
were only located in the potato cells. However, parts such as the nucleus, cell wall, and cytoplasm
were present in all specimen. Between the three power objectives, the high power objective
enabled the viewer to observe the specimens in detail. Also with the help of the iodine solution,
the images of the specimen were enhanced which made the cell easier to visualize and he parts to
be identified.

Conclusion
The purpose of this laboratory exercise was to learn how to properly prepare a wet mount
slide of different specimen such as water plant (hydrilla), potato and onion bulb. The procedure
was accomplished by a series of steps carefully executed to avoid air. This is an important skill to
develop in order to be able to study specimens with a microscope and correctly draw observations
with all proper labels.

At the cellular level, all plants previously mentioned are eukaryotic in which they have a
nucleus and other organelles and are made up of more than one cell. The cell parts in the wet mount
were not as distinct in appearance as in those of the stained specimen. Thus, the addition of stains
such as iodine solution enhanced the contrast of the images viewed under the microscope. Beyond
that, the higher the power of the microscope magnification can view more clearly the details of the
specimen.
References

Anderson, H. (n.d.). Onion cells under the microscope. Retrieved from

https://www.microscopemaster.com/onion-cells-microscope.html
Bailey, R. (2018). Learn About Plant Cell Types and Organelles. Retrieved from
https://www.thoughtco.com/what-is-a-plant-cell-373384
Deziel, C. (2018). The reason for staining a specimen on the microscope. Retrieved from
https://sciencing.com/reason-staining-specimen-microscope-5366849.html
Liu, S. (2017). Intracellular microfluid transportation in fast growing pollen tubes. Retrieved from
https://www.sciencedirect.com/topics/biochemistry-genetics-and-molecular-
biology/cytoplasmic-streaming