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MLSCI 350 > H&E / Coverslipping > Flashcards

H&E / Coverslipping Flashcards

1
Q

T or F:
H&E is performed on all tissues?

A

True

wide variety of tissue features/components

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2
Q

How many shades of pink should there be?

A

3:
light - connective tissue
med pink - muscle
dark pink/red - RBC

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3
Q

Too much hematoxylin

A

Dark purple cast over cytoplam
lack of chromatin pattern

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4
Q

Eosin Assessment

A

connective tissue - pink
muscle - med pink
RBC - dark pink/red

contrast between 2 stains

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5
Q

Hematoxylin Assessment

A

Chromatin patterns present
pink cytoplasm
dark staining lymphocyte

contrast between 2 stains

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6
Q

Hematoxyline must be: (2) prior to use

A

Oxidized into hematein to be functional
Mordanted - ferric iron or aluminum

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7
Q

purpose of a “bluing” agent

A

Alkaline solution that changes Hematoxylin from pale red to blue-black

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8
Q

Hematoxylin is a CATIONIC/ANIONIC dye

A

CATIONIC
stains negatively charged groups

phosphates - nuclear stain

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9
Q

Where do we find hematoxylin in nature

A

Logwood tree

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10
Q

eosin Y vs eosin B

A

Y - yellow
B - blue
ACIDOPHILIC dye

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11
Q

preferred pH of Eosin

A

4.0-5.9
4.5

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12
Q

Which hematoxylin demonstrates mucin

A

Gill’s

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13
Q

what is responsible for the three shades of pink?

A

Eosin being solutble (but not cleared) in water and alcohol

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14
Q

H&E maintenance

A
  • filter daily
  • rotate/replace dehydrating alcohol
  • replace stains based on SLIDE VOLUME
  • use of a control slide daily - any tissue will work
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15
Q

Poorly stained hematoxylin

poorly defined nucleus/chromatin

A
  • insufficient staining time
  • over-differeniated with acid-alcohol
  • old/expired stain
  • insufficient bluing
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16
Q

Hematoxylin stained too dark

little chromatin detail, purple cytoplasm

A
  • prolonged staining time
  • under-differentiated with acid alcohol
17
Q

Blue-purple precipitate on tissue

A
  • failure to filter hematoxylin
18
Q

Eosin pale staining

A
  • contam. with blue reagent (pH too high/alk)
  • extended dehydration
  • insufficient staining time
  • old/expired stain
19
Q

Eosin staining too intense

A
  • prolonged staining time
  • insufficient time in dehydating alcohols
  • pH too low (tech error)
20
Q

Less than 3 shades of eosin staining

A
  • insufficient time in dehydating alcohol
  • incorrect pH
21
Q

Purpose of coverslipping

A

protection from damage
focus under high magnification

22
Q

Mounting media

A

Resinous - polymers in organic solvent
Aqueous - for tissues that are soluble in organic solvents - simple syrups and preservatives

23
Q

T/F: tissue can dry before coverslipping?

A

FALSE
Drying artifact may occur

MLSCI 350 (2 decks)

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