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Microbiology and Parasitology > Module 06: Differential Staining > Flashcards

Module 06: Differential Staining Flashcards

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Biology 101 flashcards
1
Q

Punctiform are usually how large in mm diameter?

A

1mm in diameter

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2
Q

What are some common colony shapes?

A

(1) Round
(2) Filamentous
(3) Irregular

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3
Q

What are some common colony margins?

A

(1) Smooth (entire)
(2) Curled
(3) Wavy
(4) Lobate
(5) Filamentous

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4
Q

What are some colony surface characteristics?

A

(1) Smooth
(2) Concentric
(3) Wrinkled
(4) Contoured

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5
Q

What are the general forms of media?

A

(1) Broth culture
(2) Agar slant patterns

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6
Q

What are different forms of agar preparations?

A

(1) Agar slant
(2) Agar deep
(3) Agar plate

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6
Q

What are some selected broth culture properties?

A

(1) Turbidity or cloudiness
(2) Pellicle formation
(3) Sediment formation

(Note that growth is located at the bottom of the tube. The rest of the broth is clear)

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7
Q

What are some agar slant patterns?

A

(1) Filiform (even)
(2) Echinulate (pointed)
(3) Arborescent (branched)

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8
Q

What are some margin (edge) forms?

A

(1) Curled
(2) Entire
(3) Filamentous
(4) Lobate
(5) Undulate
(6) Serrate

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9
Q

What are some whole colony forms?

A

(1) Circular
(2) Granular
(3) Punctiform
(4) Filamentous
(5) Irregular
(5) Rhizoid

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10
Q

What are some surface properties?

A

(1) Concentric
(2) Contoured
(3) Radiate (ridges)
(4) Rugose (wrinkled)
(5) Smooth

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11
Q

What are some forms of elevation?

A

(1) Convex
(2) Pulvinate
(3) Umbonate
(4) Flat
(5) Raised
(6) Subsurface (as in pour plates)

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12
Q

What are some agar slant stroke cultures?

A

(1) Arborescent (branched)
(2) Beaded
(3) Echinulate (pointed)
(4) Filiform (even)
(5) Rhizoid (rootlike)
(6) Spreading

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13
Q

This type of agar slant stroke culture is characterized to be branched.

A

Arborescent

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14
Q

This type of agar slant stroke culture is characterized to be pointed.

A

Echinulate

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15
Q

This type of agar slant stroke culture is characterized to be even.

A

Filiform

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16
Q

This type of agar slant stroke culture is characterized to be rootlike.

A

Rhizoid

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17
Q

Cite the steps of differential staining.

A

(1) Before staining
(2) After primary staining (Crystal violet)
(3) After mordant, iodine
(4) After decolorizer, alcohol or acetone or alcohol
(5) After counterstain, safranin

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18
Q

Under differential staining, this is known as the primary staining.

A

Crystal violet

19
Q

Under differential staining, this is known as the mordant.

A

Iodine

20
Q

Under differential staining, this is known as the decolorizer.

A

Alcohol or acetone alcohol

21
Q

Under differential staining, this is known as the counterstain.

A

Safranin

22
Q

How should we perform differential staining?

A

(1) Most stains used in microbiology are differential stains.
(2) Use more than one dye so that different cells, chemicals or structures can be distinguished.

23
Q

How should one perform the ziehl-neelson stain kihyoun modification in differential staining?

A

(1) A small amount of an organism is suspended in a saline solution fixed on a slide.
(2) Slide is flooded using Carbol Fuschin and phenol for three (3) minutes and gently rinsed with water.
(3) Slide is decolorized with 3% HCl in70% alcohol until color appears to be removed (approx. 2 mins) and rinsed with water.
(4) Slide is flooded with methylene, which is a blue counterstain for 30 seconds, and is rinsed with water and air dried.

24
Q

This distinguishes between two large groups of microorganisms name gram positive and gram negative.

A

Gram staining

25
Q

Under gram staining, this is known as purple staining.

A

Gram positive cells

26
Q

Under gram staining, this is known as pink staining.

A

Gram negative cells

27
Q

This sis a chemical used to fix the staining reaction.

A

Mordant

28
Q

What is the difference between gram positive and gram negative cells?

A

The structure of the thinner cell walls of Gram negative bacteria cannot hold the dyes previously used, once the decolorizer is applied.

29
Q

Cite the steps of gram staining.

A

(1) Crystal violet (1 min) : rinse
(2) Iodine (1 min) : rinse
(3) Acetone Alcohol (10–15 sec) : rinse
(4) Safrinin (1 min)

30
Q

Enumerate the steps in performing gram staining.

A

(1) Cover the smear with crystal violet reagent for 1 minute. Add more stain if drying occurs.
(2) Rinse the slide (front and back) in slowly running water for 5 sec.
(3) Rinse the smear with iodine reagent and pour-off the excess. Cover the smear again with the iodine and allow it to remain for 1 min.
(4) Rinse the smear with running water as in step 02.
(5) Apply the alcohol decolorizer dropwise and slowly, Continue to add this reagent until dye no longer runs off from the smear.
(6) Rinse with running water again in step 02, to prevent additional decolorization.
(7) Cover the smear with safranin for 1 min.
(8) Rinse again with water.
(9) Blot dry but do not rub. Observe as directed.

31
Q

This form of staining is used to stain the cells of the genera.

A

Acid-Fast Stain

32
Q

These are known to cause many diseases in humans, including tuberculosis, leprosy, and other lung and skin infections.

A

(1) Mycobacterium
(2) Nocardia

33
Q

Why do mycobacterium and nocardia use acid fast staining?

A

Cells of these bacteria have large amounts of waxy lipid in their cells walls, so the Gram stain and other water-based stains don’t work well on them.

34
Q

How does gram staining work?

A

Cells are determined to be acid-fast or not, based on whether they retain their primary stain after decolorization.

35
Q

Cite the steps of acid fast staining.

A

(1) Create a smear of the organism that you are testing. Cover the smear with a strip of blotting paper.
(2) Saturate paper with Ziehl’s carbolfuchsin and heat for 3 – 5 minutes. Remove blotting paper.
(3) Wash slide with tap water and then decolorize the smear for 10 - 15 seconds with acid alcohol. Rinse again with tap water.
(4) Apply crystal violet for 1 minute, wash, blot dry.

36
Q

Enumerate the materials used for acid fast stain.

A

(1) Blotting paper : Ziehl’s carbolfuchsin (3 – 5 min heat) : rinse
(2) Acid Alcohol (10 – 15 sec) : rinse
(3) Crystal violet (1 min)

37
Q

These are characterized to be dormant, highly-resistant cells inside the cytoplasm of the bacteria, that can survive environmental extremes (desiccation, heat, harmful
chemicals)

A

Endospores

38
Q

What are some environmental extremes that endospores can survive?

A

(1) Desiccation
(2) Heat
(3) Harmful chemicals

39
Q

What are some of the most notable genera that undergo endospore staining?

A

(1) Bacillus
(2) Clostridium

40
Q

Why can’t endospores be stained?

A

Endospores cannot be stained by normal staining procedures because their walls are practically impermeable to all chemicals.

41
Q

How does endospore staining work?

A

(1) Endospore stain uses heat to drive the primary stain, malachite green into the endospore.
(2) After cooling, the sample is decolorized with water and counter stained with safranin.
(3) Results in green stained endospores and red-colored vegetative cells.

42
Q

Enumerate the materials needed for endospore staining.

A

(1) Malachite Green (5 min heat) : rinse
(2) Safrinin (20 sec)

43
Q

Enumerate the steps of bacterial smear preparation.

A

(1) Grasp the inoculating instrument with your thumb and index finger as if it were a pencil or pen.
(2) Insert the inoculating instrument at a 60 degree angle into the upper cone (hot portion) of the Bunsen burner flame. Heat the entire wire portion to redness.
(3) Pick up a tube of culture with your free hand. With broth (liquid) cultures, shake from side to side to bring the organisms into suspension.
(4) Remove the plug or cap of the tube with free fingers of the hand holding the inoculating loop.
(5) Flame the tip of the tube by passing it quickly through the upper cone of the flame.
(6) Insert the sterilized inoculating loop and remove a small amount of culture (Note: Do not be disturbed by the sizzling noise)
(7) Flame the tube again and replace the plug or cap. Put the tube in a rack.
(8a) With liquid cultures, place a loopful of broth on a slide, spread the broth over the small area, and allow the smear to dry.
(8b) With agar cultures, place a small amount of growth in a loopful of water already on the slide mix, spread over a small area and allow to air dry.
(9) Flame the inoculating instrument and put it on a metal surface
(10) Pass the smear through the hot portion of the flame, do not overheat

44
Q

This is the inoculating instrument used for liquid cultures.

A

Inoculating loop

45
Q

This is the inoculating instrument used for solid medium.

A

Inoculating Needle

46
Q

Enumerate the steps of simple staining.

A

(1) Apply only enough stain to cover the smear and allow to remain for 30 seconds.
(2) Pour off the stain. Rinse with slowly running water.
(3) Dry the slide by placing it between pieces of blotting paper. Do not rub.

Microbiology and Parasitology (9 decks)

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