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Histotechnology > H&E and Coverslipping > Flashcards

H&E and Coverslipping Flashcards

1
Q

What are the expected results of an H&E stain

A

nuclei, cartilage and microorganisms will stain blue-purple
collagen and elastin will stain pale pink
cytoplasm will stain shades of pink
muscle fiber will stain dark pink to red
RBCs and eosinophils will stain bright red

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2
Q

Where is hematoxylin from

A

it is extracted form the heartwood of the Haematoxylum campechianum tree by boiling the wood and then precipitating it by adding urea

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3
Q

What is hematein

A

the resulting dye from oxidizing hematoxylin

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4
Q

What are the two ways to oxidize hematoxylin

A

natural oxidization or chemical oxidization

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5
Q

Examples of naturally oxidized hematoxylin

A

Ehrlichs and Delafields

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6
Q

Examples of chemically oxidized hematoxylin

A

Gills, Harris and Mayers

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7
Q

What are the common mordants used in hematein

A

aluminium or iron

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8
Q

What is the proper pH range for hematoxylin

A

1.0-3.0

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9
Q

What is regressive staining

A

over-staining the section and then differentiating it with acid alcohol

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10
Q

What is progressive staining

A

no over-staining occurs, it undergoes staining for a predetermined amount of time

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11
Q

What can effect staining time

A

age of solution, formulation, progressive vs. regressive, if tissue is frozen or fixed

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12
Q

What is a disadvantage of alum hematoxylins

A

they are decolourized by acidic staining solutions

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13
Q

What are the ingredients in Gills Hematoxylin

A

distilled water
ethylene glycol
aluminum sulfate
sodium iodate
hematoxylin
acetic acid

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14
Q

What are the ingredients in Harris hematoxylin

A

distilled water
absolute ethyl alcohol
ammonium aluminum sulfate
hematoxylin
sodium iodate

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15
Q

What are the ingredients in Mayer hematoxylin

A

distilled water
sodium iodate
ammonium aluminum sulfate
hematoxylin
citric acid
chloral hydrate

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16
Q

What are the ingredients in Ehrlich hematoxylin

A

distilled water
95% alcohol
glycerol
hematoxylin
ammonium aluminum sulfate
acetic acid

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17
Q

What are the ingredients in delafield hematoxylin

A

distilled water
95% alcohol
glycerol
hematoxylin
ammonium aluminium sulphate

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18
Q

What is the less stable hematoxylin dye

A

iron hematoxylins

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19
Q

What is an example of an iron hematoxylin

A

Weigerts hematoxylin

20
Q

What is eosin

A

an acidophilic xanthene dye

21
Q

How is eosin used

A

as a 0.25-1% solution in alcohol or water

22
Q

What is the ideal pH for eosin staining

A

4.5-5

23
Q

What does the addition of phloxine to eosin do

A

produces more vivid shades of red

24
Q

What is the HPS stain

A

hematoxylin, phloxine, saffron
allows differentiation of collagen (yellow) from muscle and cytoplasm (pink)

25
Q

What is the cause of pale hematoxylin

A

insufficient stain time
old stain
over-differentiation

26
Q

What is the solution for pale hematoxylin

A

increase staining time
replace stain with fresh solution
decrease time in acid alcohol

27
Q

What is the cause of dark hematoxylin

A

too long in stain
under-differentiated

28
Q

What is the solution for dark hematoxylin

A

decrease staining time
increase time in acid-alcohol

29
Q

What is the cause of pale eosin

A

contamination of eosin with bluing reagent
extended dehydration after eosin staining
old eosin
insufficient staining time

30
Q

What is the solution for pale eosin

A

check pH
less time in dehydrating alcohols
increase staining time

31
Q

What is the cause of dark eosin

A

pH is too low
too long in eosin
insufficient time in alcohols during dehydration

32
Q

What is the solution for dark eosin

A

add NaOH to increase pH
decrease time in eosin
increase time in 70% alcohol

33
Q

What is the cause of less than 3 shades of eosin

A

pH too high
insufficient time in alcohols

34
Q

What is the solution for less than 3 shades of eosin

A

increased time in 70% alcohol
check pH

35
Q

What is the cause of a blue-black amorphous artifact overlaying the sections

A

stain precipitate from hematoxylin

36
Q

What is the solution for hematoxylin pigment

A

filter the hematoxylin before use

37
Q

What is resinous mounting media

A

synthetic resins dissolved in toluene

38
Q

What is the refractive index of resinous mounting media

A

1.51-1.55

39
Q

When is aqueous mounting media used

A

when stains or tissue elements may dissolve in alcohol or xylene

40
Q

What is aqueous mounting media

A

simple syrups, gum arabic or glycerol gelatin

41
Q

What is the refractive index of aqueous mounting media

A

1.41-1.43

42
Q

What is the cause of water droplets noted in stained sections

A

insufficient dehydration
clearing agent contaminated with water

43
Q

What is the solution for water droplets in stained sections

A

remove coverslip with xylene, return to absolute alcohol and clear with fresh xylene

44
Q

What is the cause of parts of the slide not being able to focus

A

mounting media on top of the cover slip

45
Q

What is the solution for mounting media on top of the coverslip

A

removed coverslip and remount with a clean one

46
Q

What is the cause of corn-flaking artifact

A

section allowed to dry prior to coverslipping
mounting media too thin

47
Q

What is the solution for corn-flaking artifact

A

removed coverslip, rehydrate and soak in water for 15 minutes, dehydrate, clear and remount

Histotechnology (10 decks)

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