bacterial smear preparation and simple stain Flashcards

1
Q

This is done when basic stain is applied

A

Simple staining

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2
Q

What type of stain do we use in simple staining?

A

Basic stain

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3
Q

These helps us identify the bacteria under the microscope after applying staining reagent to the prepared smear

A
  • Size
  • Shape
  • Arrangement
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4
Q

It is done after air drying the bacteria in the glass slide in smear preparation

A

Heat Fix

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5
Q

What electric charge do stain carries which the surface and cytoplasm of bacteria attracts?

A

Positive electric charge

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6
Q

What electric charge do the surface or cytoplasm of bacteria carries which stain attracts?

A

Negative electric charge

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7
Q

Two types of sample in the
DIRECT SMEAR

A
  • from SWABS
  • Concentrated centrifuged specimens
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8
Q

What do we do to avoid destruction of cellular entities in direct smear preparations when using swab?

A

Rolling the swab across the slide

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9
Q

In preparing concentrated centrifuged specimens in a glass slide, what do we use to transfer the sample?

A

Plastic transfer pipette

Needs to be sterile

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10
Q

Three types of samples for SMEARS

A
  • Culture smears (subculture smears)
  • Blood culture bottles
  • Solid media
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11
Q

How many slide do we use in preparing smears from blood culture bottles?

A

One slide only

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12
Q

It is used to puncture the top of the blood culture bottle

A

Safety venting subculture unit

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13
Q

It is used for placing a small drop of water in the squares of glass slides

(not sure with this question)

A

I-mL tuberculin syringe

with the needle removed

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14
Q

These are added first before smearing bacteria to the glass slide

A
  • Sterile NSS
  • Distilled water (H20)
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15
Q

Why do we rub the colony into the glass slide with a drop of H20 or distilled water in it?

A

for emulsification

or to emulsify

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16
Q

What should be the appearance of the glass slide after emulsifying it?

A

Turbid

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17
Q

Two types of culture

A
  • Pure culture
  • Mixed culture
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18
Q

Three types of tool used for implanting the bacteria into the glass slide

A
  • Inoculating loop
  • Inoculating needle
  • Disposable Inoculating loop and needle
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19
Q

Two sizes for inoculating loop

A
  • 10uL
  • 1uL
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20
Q

It is commonly reffered to the use of a single stain to color a bacterial organism

A

Simple staining

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21
Q

How many stain do we use in simple staining?

A

1 stain only

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22
Q

Three common types of stains in simple staining

A
  • Methylene blue
  • Basic fuchsin
  • Crystal violet.
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23
Q

Why do we use stain to color bacterial organism?

A

It is useful in determining basic morphology and the presence or absence of certain kinds of granules.

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24
Q

How many seconds/minutes is done in simple staining?

A

30 seconds to 2 minutes

depending on the affinity of the dye

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25
Q

It is where we place the glass slides for gram staining or acid-fast staining

A

Staining rack

26
Q

What are the steps in gram staining?

A
  1. Primary stain
  2. Mordant
  3. Decolorizer
  4. Secondary stain/counterstain
27
Q

Primary stain used in gram staining

A

crystal violet

28
Q

Mordant used in gram staining

A

Gram’s Iodine

29
Q

Decolorizer used in gram staining

A

Acetone alcohol

30
Q

Secondary stain/counterstain used in gram staining

A

Safranin

31
Q

Typical time for flooding a glass slide with primary stain in gram staining

A

1 minute

32
Q

Typical time for flooding a glass slide with mordant in gram staining?

A

1 minute

33
Q

Typical time for flooding a glass slide with decolorizer in gram staining

A

3 seconds or until the stain comes off

34
Q

Typical time for flooding a glass slide with secondary stain in gram staining

A

30 seconds

35
Q

What do we do between the steps performed in gram staining?

A

Wash off the slides with distilled water

36
Q

What are the steps in acid-fast staining?

A
  1. Primary stain
  2. Mordant
  3. Decolorizer
  4. Secondary stain/counterstain
37
Q

Two methods in acid-fast staining?

A

Hot and Cold method

38
Q

What does the HOT method called in acid-fast staining?

A

Ziehl-Neelsen Method

39
Q

What does the COLD method called in acid-fast staining?

A

Kinyoun Method

40
Q

Primary stain used in acid-fast staining

A

Carbol Fuchsin

41
Q

Mordant used in HOT method for acid-fast staining

A

Heat

42
Q

Decolorizer used for HOT method in acid-fast staining

A

Acid alcohol

43
Q

Counterstain used for HOT method in acid-fast staining

A

Methylene blue/Malachite green

44
Q

Typical time for flooding a glass slide with primary stain in acid-fast staining?

A

2 minutes

45
Q

Typical time for applying heat to the glass slide using alcohol lamp in acid-fast staining?

A

5 minutes

46
Q

Typical time for flooding a glass slide with decolorizer in acid-fast staining?

A

3 seconds or until the stain comes off

47
Q

Typical time for flooding a glass slide with secondary stain in acid-fast staining?

A

1 minute

48
Q

What is the color of a gram POSITIVE bacteria after applying counterstain in gram staining?

A

Purple

49
Q

What is the color of a gram POSITIVE bacteria after applying decolorizer in gram staining?

A

Purple

50
Q

What is the color of a gram NEGATIVE bacteria after applying counterstain in gram staining?

A

Pink/Red

51
Q

What is the color of a gram NEGATIVE bacteria after applying decolorizer in gram staining?

A

Colorless

52
Q

What is the color of a gram POSITIVE bacteria after applying counterstain in acid-fast staining?

A

Red

53
Q

What is the color of a gram NEGATIVE bacteria after applying counterstain in acid-fast staining?

A

Blue/Green

54
Q

What is the color of a gram POSITIVE bacteria after applying decolorizer in acid-fast staining?

A

Red

55
Q

What is the color of a gram NEGATIVE bacteria after applying decolorizer in acid-fast staining?

A

Colorless

56
Q

What is the composition of the acid alcohol used in HOT method?

A

3% hydrochloric acid (HCL) + 95% ethanol

57
Q

What is the composition of the acid alcohol used in COLD method?

A

3% sulfuric acid (H2SO4) or 0.5% hydrochloric acid (HCL) + 95% ethanol

58
Q

The position of the inoculating straight wire in the alcohol lamp

A

Diagonal downward

59
Q

Length of the inoculating straight wire

A

not more than 5cm

60
Q

Composition of the inoculating straight wire

A

Made up of: Nichrome wire (or platinum)

61
Q

We flame the inoculating straight wire until _____

A

Red hot