Microbiology Fall 2016 Study Guide Lab Exam 1

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1

What are the standard things you do upon arriving to class each day, before the activity starts?

decontaminate work surface with disinfectant

2

what are some of the standard things you do before leaving class each day?

decontaminate work surface with disinfectant

3

what needs to be written on each plate or test tube?

your name, name of organism, and date it was innoculated

4

where do we dispose of broken glass and old prepared slides?

broken glass container

5

where do we dispose of old Petri plates?

autoclaveable containers: Sharps container or orange biohazard bag

6

What are some of the objects that are regularly flamed to avoid contamination with unwanted bacteria?

inoculating loop and top of glass tube

7

what is the power of the ocular lens?

10x

8

what is the power of the four objective lenses on our microscope?

4x, 10x, 40x, 100x

9

what is the equation to calculate total magnification?

ocular lens x objective lense

10

what is the purpose of immersion oil

to increase the resolving power of a microscope

11

which objective lens requires immersion oil?

100x

12

why do we go through the effort of fusing the bacteria of the smear.

to kill the bacteria and to make sure it sticks to the slide, so when we stain, it's there

13

what are the three things we flamed during the preparation of smear

inoculating loop, top of test tube, and slide to heat fix

14

why to we flame these objects?

to avoid contamination and to make sure the bacteria stick to the slide

15

what is a wet mount?

organism on a slide in a drop of liquid, organism is still alive, requires coverslip

16

what is the advantage of using a wet mount instead of a smear?

the organisms are alive

17

why do we stain microorganisms?

some are transparent; to be able to see them in contrast

18

how is a simple stain different from a differential stain?

simple stains color the everything on the slide, differential stains color only certain types of cells

19

what are the names of the two simple stains we used in our lab?

safranin and methylene blue

20

when are simple stains used in differential staining protocol?

as a counterstain

21

what are the four basic steps of a differential stain?

  1. primary stain
  2. mordant
  3. decolorizer
  4. counterstain
22

For gram staining what is the primary stain ?

crystal violet

23

For gram staining what is the mordant

gram's iodine

24

For gram staining what is the decolorizer?

acetone or alcohol

25

For gram staining what is the counterstain?

safranin

26

For gram staining which of the chemicals used in the staining procedure that we did in lab does not match the lab book?

alcohol/acetone

27

what color is gram positive cells at the end of the stain?

purple

28

what color is the gram negative cells at the end of the stain?

pink

29

what is the structural difference between gram - and gram + cells?

gram - has two plasma membrane and one thin cell wall

gram + have a thick cell wall

30

what shape and color is the bacteria of Staphylococcus aureus

cocci, purple

31

what shape and color is the bacteria of Bacillus brevis

bacili, purple

32

what shape and color is the bacteria of Escherichia coli

bacili, pink

33
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Gram + and Gram - staining

34

For spore staining, what is the primary stain?

malachite green

35

For spore staining, what is the mordant

steam

36

For spore staining, what is the decolorizer?

water

37

For spore staining, what was the counterstain ?

safranin

38

For spore staining, what are the two general of common spore formers?

Bacillus and Clostridium

39

Where do spore-forming bacteria commonly grow?

survive environmental conditions that are not favorable

40

what color are the endospore at the end of the stain?

green

41
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Spore staining

42

why do bacteria form endospore?

spore are produced when conditions become unfavorable

43

what appear pink at the end of the spore stain?

the vegetative cells

44

For acid-fast staining what is the primary stain ?

basic fuchsin

45

For acid-fast staining what is the mordant

steam

46

For acid-fast staining what is the decolorizer?

acid alcohol

47

For acid-fast staining what is the counterstain?

methylene blue

48

For acid-fast staining what chemical was used for the primary stain in our lab ?

carbol fuchsin

49

What color are the acid fast positive cells at the end of the stain?

metallic shiny red

50

what is the genus of the organism that was used as a acid-fast negative control?

Staphylococcus

51
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Acid-fast staining

52

For capsule staining how is the basis of a negative stain different from the other staining protocols ?

The background is dark and the cells are clear.

53

how is the fixing step for the capsule stain different from the other staining protocols that we performed so far in lab?

it is chemically fixed with acid alcohol. and not heat fixed

54

For capsule staining what is the primary stain?

congo red stain

55

For capsule staining what is the purpose of acid alcohol?

fixing the smear on the slide

56

For capsule staining what is the secondary stain ?

carbol fuchsin

57

For capsule staining what is the genus of the organism that was used in class for this stain?

Klebsiella

58

For capsule staining what color is the capsule at the end of the procedure?

mostly clear or colorless

59

For capsule staining what color is the background?

purple/grey/red

60

how does the presence of a capsule affect the pathogenicity of a bacterium?

the capsule shield from immune response

61
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Capsule Staining