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350 > Coverslipping > Flashcards

Coverslipping Flashcards

1
Q

T/F hematoxylin is a dye

A

F - it must be oxidized to form hematein which is the dye

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2
Q

how does chemical oxidation work

A

using sodium iodate to convert hematoxylin to hematein

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3
Q

does natural or chemical oxidation have a longer shelf life

A

natural

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4
Q

why are all hematoxylin used at low pH

A

suppress non nuclear staining

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5
Q

what pH is hematoxylin

A

pH ~ 2.5

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6
Q

what color does alum hematoxylin stain nucleus

A

brick red to purple when washed in alkaline solution (bluing)

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7
Q

what are good bluing agents

A

lithium carbonate, dilute ammonia, Scott’s tap watear

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8
Q

what is a disadvantage of alum hematoxylin

A

decolorize by acidic soultions = cannot do masson trichrome

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9
Q

special ingredients in Gill hematoxylin

A

aluminum sulfate mordant
ethylene glycol solvent

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10
Q

what is significant about Gill hematoxylin

A

alcohol free and stains goblet cells

progressive stain

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11
Q

special ingredients in Harris hematoxylin

A

absolute ethyl alcohol solvent

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12
Q

what is significant about Harris hematoxylin

A

used to be used with mercury

regressive stain

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13
Q

special ingredients in Mayers hematoxylin

A

citric acid
chloral hydrate preservative

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14
Q

what is significant about Mayers hematoxylin

A

alcohol free

progressive OR regressive

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15
Q

special ingretiants in Erlich hematoxylin

A

glycerol preservative
acetic acid

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16
Q

what is significant about Erlich hematoxylin

A

naturally oxidized

regressively

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17
Q

special ingredients in Delafield hematoxylin

A

glycerol presrvative

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18
Q

what is significant about Delafield hematoxylin

A

naturally oxidized

regressively

19
Q

what is iron used as in iron hematoxylin

A

mordant and oxidizer

20
Q

what is the most common iron hematoxylin

A

weigerts hematoxylin

21
Q

how is weigerts hematoxylin prepared

A

fresh using solution A and B

A- hematoxylin
B - iron with HCL

22
Q

difference in staining between iron and alum hematoxylin

A

iron can only be use PROgressivly

alum is for both

23
Q

disadvantage of iron hematoxylin

A

time consuming set up and prolonged staining

24
Q

What is thymol used for

A

control microbes in Eosin

25
Q

what is used to adjust pH of eosin

A

acetic acid

26
Q

what pH should eosin be at for best staining

A

4.5- 5

27
Q

what does phloxine B and eosin do

A

more vivid shades of red

28
Q

what does an HPS (hematoxylin, phloxine, saffron) stain do

A

differentiating yellow collagen from muscle and cytoplasm (red)

29
Q

where can sections be lost during staining

A

bluing if agitated

30
Q

cause for pale hematoxylin

A

less time
old stain
over differentiated

31
Q

cause of dark hematoxylin

A

too long
under diff

32
Q

cause for pale eosin

A

pH too high from bluing agent - add acetic acid
less time
prolonged dehydration

33
Q

cause for dark eosin

A

pH too low - add NaOH
too long
too low time in dehydration

34
Q

less than 3 shades of red

A

too low time in dehydration

35
Q

artifact

A

did not filter hematoxylin

36
Q

purpose of coverslip

A

presrve stained tissue, protect, focus at high powers

37
Q

what is the refractive index of resinous mounting media

A

1.5

38
Q

when is aqueous mounting media used

A

when fat in tissue will be dissolved in xylene

39
Q

disadvantage of aqueous mounting media

A

low refractive index = less focus on high power
not permanant
bleeding of stains

40
Q

what helps aqueous mounting media from bleeding

A

adding sugars

41
Q

what happens when a slide dries before coverslipping

A

corn flaking = forms brown articfact

42
Q

cause of water droplet on stained s ection

A

low time dehydrating
clearing agent (xylene) contaminated w water

43
Q

cause of unfocussed parts of slide

A

mounting media on top

350 (7 decks)

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