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Lab Test 1 Flashcards

1
Q

What is the Kinyoun Stain used for

A

Acid-fast species of the bacterial genus Mycobacterium. It is a variation of a method developed by Robert Koch in 1882

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2
Q

Which was the Mordant used in the Acid-fast stain

A

DMSO

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3
Q

In our lab what experiment did we use: M. Smegmatis & S. epidermidis

What were the results

Describe appearance of each

A

Acid-fast stain

Staph epidermis = Not acid fast (Cocci Blue)

Mycobacterium smegmatis = Acid fast. (Pink Bacillus)

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4
Q

Why is DMSO used in fast-acid staining?

Which stain does it allow to penetrante & attach to the Waxy Cell Wall?

A

It is a Surfacent and allows Carbol Fuchsin to penetrante the Waxy cell wall

Carbol Fuchsin

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5
Q

Which type of bacteria are associated with Acid-fast cells

Why?

A

Mycobacteria

They have a Waxy Cell Wall

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6
Q

A Positive indication for Acid-Fast bacteria is….

A negative is

A

Positive = Red / Pink

Negative = Blue

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7
Q

Name the alcohol used in Acid Fast staining

Name the alcohol used in gram staining

A

Acid fast = Acid Alcohol

Gram staining = Aceton Alcohol

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8
Q

2 stains used in Acid Fast in Order

Are they basic or acidic

Positive or Negative charged

Reagent

Decolorizer

A

Carbol Fuchsin

Methylene Blue

Basic

DSMO

Acid Alcohol

Positive charged

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9
Q

How long does the DMSO-carbol fuchsin stay on the slide

A

4 minutes

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10
Q

How many passes with the Acid Alcohol in the Acid-Fast decolorizing technique

How many passes with the Aceton Alcohol in the Gram-staining Tech

A

1 pass

2 passes

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11
Q

The fast Acid cells are described as “pleomorphic”

True or False

A

True

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12
Q

When do you blot slides?

A

At the very end

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13
Q

Name the reagents used for gram-staining in order.

How long do you leave each on for?

A
  1. Crystal Violet 1 min (Primary Stain)
  2. Grams (Lugols) Iodine 1 minute (Mordant)
  3. Acetone alcohol (Decolorizer) Remove instantly
  4. Safranin 1 minute (Counter Stain)
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14
Q

Decolorizer, decoloizes….

A

Gram-negative & non acid fast

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15
Q

In the Gram Staining experiment which 2 bacteria were looked at?

What were the results?

A

Staphylococcus epidermidis = Gram Positive (Purple Grapes)

Escherichia coli = Gram Negative (Red Rods)

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16
Q

Name of paper for drying slide

A

Bibulous paper

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17
Q

Wet mount 4 steps

A
  1. Loopful of culture
  2. 1 drop of stain
  3. Cover slip
  4. View with 4, 10, 40x
    NO Oil Immersion
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18
Q

Difference between Iris Diaphragm & Condensor lens

A

Iris Allows more light

Condenser Focuses it

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19
Q

What does parfocal mean

A

Ability to switch magnification without readjusting

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20
Q

2 physical characteristics of Corynebacterium

A

Rod shapped bacillus

Palisades arrangements

Gram-positive

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21
Q

Staphylococcus is gram-negative or gram-positive?

Describe shape and arrangement/ color after gram stain

A

Gram-positive

Round clusters like grapes

Purple

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22
Q

What are chromophores

A

Color part of stain

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23
Q

Difference between basic and acidic dyes

A

Basic dyes have a positive charge and adhere to the cell (due to the cell having a negative charge. )

Acidic dyes have a negative charge and stain the background (allowing the cell to be seen)

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24
Q

Crystal Violet is an example of what kind of stain?

A

Simple stain

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25
Where are fusiform bacteria found in humans
Mouth
26
Can you ID bacterial species using a Simple Stained slide?
No, simple stain only IDs the morphology
27
How do we store and label petri dishes? Why?
Label and store "upside-down" To prevent condensation
28
Disinfectants are effective against which type of pathogens? Which are they not effective against?
Vegetative cells & viruses Not effective against ENDOSPORES
29
Which is our disinfectant in the laboratory?
Vesphene
30
This type of medium allows for growing of isolated colonies and are easy to observe Disadvantage: they can dry out easily
Agar plates
31
These allow for growing large number of bacteria and at varying oxygen levels Disadvantage: you cannot isolate them
Broths
32
These dry out less easily than agar palates and take up less space. But have less visibility than agar plates
Slants
33
Retrieving a culture from a tube with a loop, the cap of the tube is removed and held with the fingers from which hand?
The one with the loop
34
Kinyoun Method is for what
Acid fast / Mycobacterium
35
The Schaeffer-Fulton method is also called...
(Endo)Spore Stain
36
How long do we let the Methylene Blue stay on the Kinyoun Method?
30 seconds
37
The endospore species is... Where is it found? Aerobic ? Gram-negative or gram-positive Morphology and arrangements
Bacillus Cereus Soil/ Yes Gram-positive Streptobacillis
38
After we air-dry and heat fix the endospore slide we add Malachite Green and then
Set on heated hot plate for 5 Minutes
39
How long do we let the malachite green cool after we take it off the 2nd hot plate?
3 minutes
40
What is the stain used in "negative stain"
Nigrosin
41
With the negative stain what were the 2 species we examined? What was the alternative choice
B. Cereus & S. Epidermidis Mouth biofilm
42
What steps do we skip in both the negative stain and the capsule stain?
Heat fixing Rinsing with distilled water
43
Which is species is in the purple broth? Which staining method is it used for? Gram positive or negative Morphology and arrangements
Enterobacter aerogenes / Klebsiella aerogenes Capsule Stain Gram negative Diplobacillus
44
Which stain do we use Aqueous Crystal Violet for? 1 minute
Capsule stain
45
In the capsule stain we use which three reagents and for how long?
Nigrosin Aqueous Crystal Violet 1 minutos 20% Copper Sulfate 3 minutos
46
What do we use to rinse the slide in the capsule stain?
20% Copper sulfate
47
Acid fast bacteria are also (gram-negative/ gram-positive)
Gram-positive
48
Name the 2 reagents used in endospore staining, in order When do we add them Do we rinse, if so with what?
after air dry and heat fixing, place on hot plate and add Malachite Green 5 minutes Remove from hot plate and rinse with distilled water Safranin Red & Rinse with distilled water
49
Kinyoun method aka Reagents Mordant Bacteria we looked at, results Color
Acid-fast Basic fuchsin 2ndary stain Methylene Blue DSMO Mycobacterium smegmatis = Acid fast Red S. Epidermidis = non acid fast Blue
50
What are the colors and the species from the Schaeffer-Fulton method. Name of species used
Red = vegetative cells Green = endospores Bacillus cereus
51
Exosporium does what what is it associated with? Composition
Protective barrier for endo spore. Proteins
52
What is the Mordant in the endospore staining
Heat
53
What is the 2ndary stain in the Schaeffer-Fulton endospore stain?
Safranin
54
Which species doesn't produce endospores Clostridium, Mycobacterium, Bacillus
Mycobacterium
55
Mycobacterium tuberculosis is visible under which kind of staining? Results from this stain
Kinyoun Acid-fast Pink
56
Advantage of Kinyoun staining over Ziehl-Neelsen? Which did we use?
Kinyoun doesn't involve heating releasing poison gas Kinyoun
57
Name acid-fast genuses
Mycobacterium & Nocardia
58
What makes Mycobacterium resistant to staining
Mycolic acid
59
Describe E coli Gram status Morphology and arrangements
Negative (Pink) Bacilli Single or Diplobacillus
60
Staphylococcus epidermidis before primary stain
Colorless
61
Pseudomonas fluorescens after primary stain?
Purple
62
Pseudomonas fluorescens Gram status Morphology and arrangements
Negative Bacillus Single
63
Bacillus megaterium after the addition of the Mordant
Purple
64
Bacillus megaterium Gram status Morphology and arrangements
Positive Streptobacillus
65
Staphylococcus epidermidis after Decolorizer
Purple
66
Moraxella catarrhalis after Decolorizer
Colorless
67
Moraxella catarrhalis Gram status Morphology and arrangements
Negative Diplococci
68
Pseudomonas fluorescens after Decolorizer
Clear
69
Bacillus megaterium after counter stain
Purple
70
Escherichia coli if you forget to add the counter stain
Clear
71
Escherichia coli if you forget to apply the Decolorizer
Purple
72
Bacillus megaterium if you forget to add Mordant (Grams Iodine)
Red
73
On the pipette pump where is 1ml?
At the 0 mark
74
0.1 ml is how much on the pipette pump
Between any 2 whole number lines
75
Standard plate count 1 ml from each test tube to the next filled with 9 ml What is the dilution of A B C
A = 1: 100 B = 1: 10,000 C= 1: 1,000,000
76
1 ml from test tube B on the plate. What is the total dilution?
1:10,000
77
0.1 ml from testtube B on the plate has What total dilution?
1:100,000
78
1 ml from testtube C on the plate has what total dilution?
1:1,000,000
79
0.1 ml from test tube C has What total dilution?
1:10,000,000
80
What is the control in the standard plate count?
1 ml E. Coli
81
Simple staining is...
Using 1 dye to stain a bacterial smear
82
Type of attraction between cell and stain
Electrostatic
83
Describe shape of Fusiform bacteria Which group of cell shapes do they fall into?
Bacillus But with tapered ends
84
Staphylococcus aureus, Streptococcus lactis, Bacillus megaterium and Corynebacterium xerosis were examined by this staining type
Simple
85
Pleomorphism refers to
Demonstrating several different shapes
86
Distinct reddish purple granules with in a cell that show up with Methylene Blue
Metachromatic granules
87
3 things you can discern with a good smear
1. Morphology (Rods, Cocci, Shapes) 2. Arrangement ( Single, Stapho, Strepto) 3. Internal structures ( endospores & cell inclusions)
88
Goals of a good smear
1. Adhere cells so they are not washed off during the staining and rinsing 2. Ensure shrinkage doesn't happen to cells. ( causing Distortion & Artifacts) 3. Prepare thin smears
89
For solid media when creating a smear which do you add first the smear or the water?
Water
90
Where do we write the name of the organism on a slide
Left-hand side
91
A hot loop will cause what to happen to the bacteria
They will go into the air
92
When do we flame the tube?
After opening After putting the inoculation loop in
93
Protoslo does...
Slows movement of bacteria
94
Gram staining is which kind
Differential

Pathogenic Microbiology (31 decks)

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