Lab assignment 1: Microbiology and microorganisms

Answer the ques,ons directly in the space provided. Do not a8ach addi,onal pages.
By entering your name below, you cer,fy that you have completed the lab yourself, and the answers
given are your own. Copying assignments of other students, or other forms of plagiarism will result in a
grade of zero, and will be reported to the Faculty of Science.

Name: ____Angela Aluloska __________________ Student number: ___007914222__________

When your assignment is complete, submit it to the assignment 1 drop box on UM Learn. This
assignment is due at the end of Lab 1.

Exercise 1: Ge,ng to know your microscope

1) How should you posi,on the condenser on the Nikon E100 bright-field microscope? What does the
condenser do? (1 mark)
When using a Nikon E100 bright-field microscope it is necessary for the condenser to be posi,oned as
close to the stage as possible for the best illumina,on. The condenser is a large lens that is used to focus
the light coming from the light source below into a narrow beam to illuminate the specimen on the slide.
2) How should you posi,on the iris diaphragm when viewing a coloured specimen, like stained
bacteria? Explain why. (1 marks)
The slider should be posi,oned to the leZ, so that the iris diaphragm is wide open to allow full light to
pass through the condenser onto the specimen. In order to see coloured specimen in the best resolu,on
possible, the iris must be en,rely open to provide excellent illumina,on.
3) When should immersion oil be used? Explain why we need immersion oil. (2 marks)

Immersion oil should always and only be used with the 100x objec,ve. Immersion oil has the same
refrac,ve index as glass, which mean that it prevents the bending of light that would occur if the light
were passing through air. Therefore the oil helps to make sure that all light passing through the specimen
reaches the objec,ve, giving the best resolu,on possible.

4) The objec,ves on our laboratory microscopes are parfocal. What does that mean? (1 mark)
Parfocal means that when one object lens is in focus, then the other objec,ves will also be in focus. Only
small adjustments with the fine focus knob are necessary.
 Explain, in your own words, how you should go to the 100x objec,ve, aZer you’ve already found
your specimen with the 10x objec,ve. (1 mark)
If I have already detected my specimen with the 10x objec,ve I can switch directly to the 100x
objec,ve. Since the objec,ves are parfocal, my specimen will not be lost when I switch to a higher
powered objec,ve. To make small adjustments to the focus, I will use the fine focus knob. The
course focus knob should not be used on higher powered objec,ves because the specimen will be
lost from view.

Exercise 2: How can you tell the difference between prokaryotes and
eukaryotes

5) Examine the picture showing the mixed bacteria (3 types) slide in the online Lab 1. What three cell
shapes are shown on this slide? (1 mark)
Bacilli (rods), cocci (spheres) and spirilla (spirals) are three cell shapes visible on the slide.

6) Explain why we almost always use the 100x objec,ve when viewing bacteria with the microscope in
the lab (rather than using lower powered objec,ves, like the 40x). (1 mark)
100x objec,ve provides the highest magnifica,on, which is necessary because bacteria (prokaryotes) are
incredibly small. With lower magnifica,on such as 40x or 10x, we would not be able to view them.

7) Examine the picture of the Halobacterium salinarum slide. What shape are the cells of
Halobacterium salinarum? (0.5 marks)
The cells are bacilli (rod) shaped.

8) How does the cell size of Archaea compare to that of the cells in the mixed bacteria (3 types) slide?
(Look carefully at all of the bacterial cells, including the largest cells on the slide.) Can you tell the
difference between Archaea and Bacteria by size? (1 mark)
Archaea and bacteria resemble each other in shape and size. The bacilli cell shapes of the Archaea
are very similar in size to the spirilla and some bacilli cell shapes of the bacteria.
9) Examine the picture of Anabaena. Are any special structures shown? If so, which one(s)? (1 mark)
Yes, there is a special structure shown on the Anabaena called a heterocyst.

10) Is Anabaena a prokaryote or a eukaryote? (0.5 marks)

Anabaena is a prokaryote.
11) Examine the short video of Euglena. In the cells in the video there is a red spot clearly visible. What
is this red spot? (0.5 marks)
The red spot is the s,gma, a structure that allows Euglena to sense light.
12) Examine the picture of Saccharomyces cerevisiae. How was the iris diaphragm posi,oned to take this
picture? Explain why. (1 mark)
Since the picture of the Saccaromyces cerevisiae is colourless, closing the iris diaphragm par,ally will
restrict the amount of light passing through the condenser and give contrast between the colourless
microbes and the colourless background.

13) How do the cells of S. cerevisiae compare to the cells of prokaryotes that you observed on previous
slides? (1 mark)
Compared to the prokaryo,c cells observed previously, the cells of S. Cerevisiae have internal
organelles.

14) Examine the picture of Penicillium. What are the structures shown at the end of the branched
hyphae? (0.5 marks)
The fungal reproduc,ve structures called conidia.

Exercise 3: Where can I find a good bacterium when I need one?

15) Describe the characteris,cs of the following colonies using the terms on pages 18-19 of the lab
manual. Make sure you are describing the colonies labeled on the pictures with red arrows. (4
marks)

Colony #1: Describe the colony indicated with the le:er A on the plate le= open to the Air.
• Form: circular
• Margin: undulate
• Color: red and white (pinkish)
• Surface quali,es: glossy
 Do you think this colony is a bacterium or a fungus?
Bacterium
Explain why, based on the features of the colony:
Doesn’t have the ‘fuzzy’ appearance as fungal colonies and is also super small.

Colony #2: Describe the colony indicated with the le:er B on the second ”washed and unwashed
fingers” plate.
• Form: circular
• Margin: undulate
• Color: grey
• Surface quali,es: dull
Do you think this colony is a bacterium or a fungus?
Fungus
Explain why, based on the features of the colony:
Has a ‘fuzzy’ appearance because of the hyphae and is rela,vely larger in size.

16) Give two ,ps to avoid contamina,on of your plates in future lab periods. (1 mark)

Tip 1: Try not to breathe into your plates- work away from your body and close to the Bunsen burner

Tip 2: Make sure you always wipe down your bench with disinfectant before you start (and before
you leave)

17) Examine the “Air” plate on the online Lab 1 results page. At what temperature was this plate
incubated, and for how long? (1 mark)
It was incubated in a 28°C incubator for 7 days.
18) How do you think it’s appearance of the ‘Air’ plate would have changed if it was incubated at 37°C
for 24 hours? Consider what you know about the growth of both bacteria and fungi, and describe
how the colonies of each would differ if the incuba,on condi,ons were changed.

How would the colonies of bacteria differ if the plate was incubated at 37°C for 24 hours? (1 mark)

The colonies of bacteria would grow very quickly at 37°C, showing growth of visible colonies in about
16 hours.

How would the colonies of fungi differ if the plate was incubated at 37°C for 24 hours? (1 mark)

Fungi oZen grow best at a temperature of 28°C, therefore at 37°C fungi will grow much slower.

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