INTRODUCTION

A microscope is an instrument widely to magnify and resolve the image of an object that
is otherwise invisible to naked eye. Wet mount method allow for observation of living
organisms. The wet mount tends to dry out quickly under the heat of the microscope light; it is
simpler to perform than the wet mount, but it is useful for short-term observation only. If there is
reason to suspect that a food has caused food poisoning or has undergone microbial spoilage, the
original product or a low serial dilution of it should be used to prepare a slide for direct
microscopic examination. A microscopic examination must be made, even though the
food may have undergone heat treatment and the microorganisms involved may no
longer be viable. To understand this principle, an experiment was being conducted at Lab 14B.

OBJECTIVES

a) To familiarize with the pertinent features of the compound microscope.

b) To understand its components and functions.
c) To familiarize with microscopic examination of microbes using wet mount method.

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RESULTS

Microorganism: Lactobacillus acidophilus (Bacteria)

Magnification : 40X10

Microorganism: Saccharomyces cerevisiae (Yeast)

Magnification : 40X10

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Microorganism : Aspergillus niger (Mould)

Magnification : 40X10

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DISCUSSIONS

This experiment was about microscopic examination of food and bacterial staining. We had used
binocular microscope as a medium to enlarge the image of bacteria, yeast and mould that used in
the experiment such as Lactobacillus acidophilus, Saccharomyces cerevisiae and Aspergillus
niger. In this experiment we have learned about the components and functions of microscope.
The main component of microscope is an eyepiece/ocular lens that is the top of lenses usually
have 10X power. It can be adjusted to suit with the user’s eye position. There are 4 objective
lenses on the microscope, where consist of 5X, 10X, 40X and 100X of power. Other than that,
the main components of microscope are light source, condenser, iris diaphragm, base and stage.
Each of the main components has their important for the usage of microscope in order to get the
best image of microbe.

Figure 4.1 Binocular microscope

For the preparation of microscopic slide of culture for examination, we have doing the
bacterial culture first. Bacterial culture is the step to make the heat-fixed smear on a slide. After
doing the bacterial culture, the slide is ready for staining purpose and microscopic observation.
Gram staining is a bacteriological laboratory technique used to differentiate bacterial species into
two large groups such as gram-positive and gram-negative based on the physical properties of

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their cell walls. Before observing the bacteria, we need to put immersion oil on the slide as
conventional mounting medium as it helps to obtain clear images of Gram-stained bacteria.

“The Gram stain has become the most important staining procedure in the bacteriology
laboratory, because it differentiates between “Gram-positive” and “Gram-negative”.” (Burton’s
Microbiology for the Health Sciences by Paul G. Engelkirk and Janet L. Duben-Engelkirk, 9th
Ed. 2011)

The results show the bacteria of Lactobacillus sp. is a gram-positive because under
microscope the colour of cell wall is purple/violet. Stain dark purple due to retaining the primary
dye called Crystal Violet in the cell wall. It has a single membrane that surrounded by a thick
peptidoglycan. The shape of Lactobacillus sp. is rod-shaped and we are used 40X10
magnification power to examine the image of bacteria. Besides that, for the Saccharomyces sp.,
the result shows it is a gram-negative because based on our observation under microscope the
colour of cell wall is red. This red colour is due to retaining the counter staining dye called
Safranin. The shape of Aspergillus niger is a circle with filamentous extensions growing out
from it and we used 40X10 magnification power to examine the image of mould.

Wet mount is a procedure for the yeast and mould culture. It is made to observe under
microscope. There are few steps to follow before observation whereas drop a water on a clean
piece of slide. Then transfer the culture into slide and mixed it into the drop of water until a
suspension is achieved. The smear of culture on flame to heat fixed the bacteria on slide. The
result shows the shape of yeast under 40X10 magnification power is like rounded shape that has
a strain. Based on observation, there a lot of cultures under a single of slide because the yeast
undergoes two reproductions where are fission yeast and budding yeast. For the mould
(Aspergillus niger), the result shows that only spores can be seen under 40X10 magnification
power because maybe while doing the bacterial culture, there are some error on taking the
specimen into the slide. The mould and yeast are eukaryotic microorganisms but the yeast is used
for the food production while the mould is a fungus that gives contaminant on food.

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CONCLUSION

Binocular microscope was used to enlarge the image of microorganisms. By using this
device, it easier to identify which category these microorganisms belongs to and it also help us to
identify the shape of these microorganisms. Each lens of the microscope has different power.
The eyepiece lens power remains constant while the four objective lens have different power.
The higher the lens power, the nearer the image was magnified by the lens. Other than that, the
light source, condenser, iris diaphragm, base and stage also has an important roles during
examining the microorganisms. Before using the microscope, the samples or microorganisms
need to be prepared. The method use during this experiment is wet method. Other than that, by
using the wet method, Gram stain positive and Gram stain negative was able to be identified
between these three microorganisms. During the experiment, there was a mistake during the
sample preparation, where the bacteria was unable discovered through the microscope, so the
sample need to be redone. Overall, the objective was achieved.

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REFERENCES

1) http://education-portal.com/academy/lesson/the-gram-stain-theory-and-
procedure.html#lesson
2) Biotechnology and Biomedical Engineering : Amrita Vishwa Vidyapeetham Virtual Lab.
(n.d.). Retrieved September 28, 2014.
3) Anderson, H. (n.d.). How does a Compound Microscope Work? Retrieved from
http://www.microscopemaster.com/parts-of-a-compound-microscope.html
4) Kalab, M. (2009, February 2). FOODS UNDER THE MICROSCOPE. Retrieved from
http://www.magma.ca/~scimat/foodmicr.htm

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