MIC60104 Tutorial 1

2020
1. List 3 domains of microorganisms.
Bacteria
Archaea
Eukaryotes

2. Which type of microscope is best for observing double-stranded DNA virus?

Electron microscope

3. Which type of microscope is best for observing motility of microorganism?

Dark field

4. Which type of microscope is best for observing internal inclusion body of a microorganism?
Phase contrast

5. To increase the contrast of microbial cells viewed under a light microscope, what would you do?
Perform staining

6. In viable cell count technique, the plating volume for spread plate method is
and for pour plates method.

100ul and 1000ul

7. What is the function of positive staining? How does it work?

Stained bacterial cells with basic dye against unstained

background
8. What is the function of negative staining? How does it work?

Unstained bacterial cells against stained background with

acidic dye (negative charged)

9. Special staining is used to enhance visibility of which part of microorganism?

to enhance features of specific part of specimen

10. What types of specimens should be chemically fixed as opposed to heat-fixed?

Tissue specimens

11. What staining method is used to differentiate cell type based on the thickness of peptidoglycan
in cell wall?
Gram stain

12. What staining method is used to differentiate cell type based on the presence of mycolic acid?
Ziehl-Neelson Staining

13. Explain the role of Gram’s iodine in the Gram stain procedure.
A mordant is a substance used to set or stabilize stains or dyes; in this case, Gram’s iodine acts
like a trapping agent that complexes with the crystal violet, making the crystal violet–iodine
complex clump and stay contained in thick layers of peptidoglycan in the cell walls.
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14. Explain the role of alcohol in the Gram stain procedure.

decolorizing agent

15. Explain the difference between simple and differential staining

In Simple staining, a single dye is used to emphasize particular structures in the specimen. A
simple stain will generally make all of the organisms in a sample appear to be the same color,
even if the sample contains more than one type of organism

In differential staining, it will distinguish organisms based on their interaction with multiple stains.
Two organisms in a differentially stained sample may appear to be different colors.

16. Name the group of microorganisms that can be found mostly in extreme environment.
Archaea
17. When the cells of Archaea are stained with Gram stain, the cells appeared as pink cells when
observed under a microscope. Why are the cells appeared as pink cells?
No peptidoglycan, consist of S-layer

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18. Name and describe the function for each of the labelled structures shown in figure below.

M: Outer membrane- Outer membrane proteins-target site for antibiotics.

N: Lipopolysaccharide- This layer consists of lipid A to which is attached a
polysaccharide
O: Membrane protein- connects the peptidoglycan to outer membrane.
P: Periplasmic space- Space between inner membrane and outer membrane.
Q: Peptidoglycan- polymer consisting of sugars and amino acids that forms a mesh – like
layer outside the cell membrane of most bacteria forming cell wall.

19. The apparatus shown in figure below is used in microbiology laboratory

for .

Streaking

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 MIC60104 Tutorial 1
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20. The method used for isolation of microbial colonies shown in the figure below is
called

Pour plate

21. Draw the following shape and arrangement of bacterial cells.

• coccus
• Streptococcus sp.
• Staphylococcus sp.
• Bacillus sp.

22. Based on the serial dilution and plate count below shown in figure below, calculate the estimate
number of cells of E. coli culture, in the expression of colony forming unit/ ml.

E. coli culture

TNTC – too numerous to count

14 x 107

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