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Histology2 > final review > Flashcards

final review Flashcards

1
Q

whats orthochromic

A

orthochromic is when tissue stains the same color as the dye

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2
Q

whats leuco compound

A

in schiff reagent when the quinoid ring loses it color it’s called leuco compund which is colorless

then reacts with aldehydes to form a new color

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3
Q

why do we hydrate tissues

A

to remove parrafin & make tissue more miscible with stain ( stain is water based)

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4
Q

when would you not hydrate tissue

A

when stain is alcoholic stop at 70 %

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5
Q

why do we dehydrate

A

to prepare for resinous mounting media

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6
Q

when would you not dehydrate

A

when using a water based mounting medium
ex, in oil red O we use glycerol as mounting media

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7
Q

do you DCM frozen sections

A

we DCM frozen sections, we dont hydrate( bc no parrafin) but we still need to mount

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8
Q

how to ripen hematoxylin

A

naturally : sunlight & dela field
artificailly : sodium iodine

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9
Q

aluminium mordant vs iron mordant

A

aluminum ( preferred) - longer shelf life

iron mordant - shorter shelf life

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10
Q

Accentuators

A
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11
Q

what does chloral hydrate do

A

prevents scum in hematoxylin( don’t need to filter)

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12
Q

why is bluing so important

A

goes from red ( soluble ) to blue ( insoluble )
nuclear stain is less likely to be removed

ex. scotts tap water = bluing agent

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13
Q

H&E - white spots following deparaffinization

A

clearing agent + water = cloudy or white spots

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14
Q

nuclear stain not crisp

A

fixation not complete

oveheated

poor processing

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15
Q

Pale nuclei

A

not enough dye or poor quality dye in the section

not enough section in dye

overexposure to acid

thin section

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16
Q

overstained or dark nuceli

A

too much dye in section

too much section in dye ( too thick)

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17
Q

red or brown nuclei

A

over oxidized hematoxylin

not enough bluing

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18
Q

pale cytoplasmic staining

A

pH

not enough dye in section

not enough section in dye

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19
Q

cytoplasm overstained

A

too much dye
too much section

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20
Q

eosin not 3 different shades

A

fixation

dehydrate & clear well

70% best for differentiation

pH 4.6-5 recommended

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21
Q

blue - black precipitate on top

A

hematoxylin was not filtered & metallic sheen was picked up on sections

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22
Q

hazy or milky water on slides

A

clearing agent & water have come together

not miscible

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23
Q

water bubbles seen in microscopically

A

incomplete dehydration

remove coverslip with xylene ( in chemical hood ), dehydrate, return to xylene, coverslip

change solutions of clearite 3 and alcohol

check for hazy milky slides before mounting ( saves time )

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24
Q

uneven H& E staining

A

contamination of wax in closed processor with water or fiaxative

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25
Q

dark staining on edge

A

laser or electrocautery

cannot fix burned tissue

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26
Q

poor contrast nuclei vs cytoplasm

A

poor nuclear staining

excessive cytoplasmic staining

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27
Q

good technique for H&E

A

checking slides after each staining step

do not allow sections to dry during procedure ( gloossy black nuceli & brown stippling)

keep solutions covered when not in use

28
Q

how to make & store schiff reagent

A

treat basic fuchsin( parosaniline) with surlfuric acid ( breaks quinoid ring ) to form colorless compund ( leucofuchsin)

combine with aldehyde will form color bright red) only after WASHING to remove surfuric acid

store in fridge but bring to room temp before use

test with formaldehyde bc aldehydes form with schiff reagent

smells like sulfur bc of surluric acid

29
Q

another name for PAS

A

McManus method

30
Q

how to make PAS more specific for glycogen

what fixative is preferred for glycogen

A

amylase or diastase = more speciific

alcohol prevents glycogen streaming

31
Q

whats a good control for glycogen

A
  • liver
  • endo/ecto cervix
32
Q

whats a good control for basement membrane

A

kidney

33
Q

metachromatic stains

A

stain of tissue is a different color than the original color of dye

change of color is due to polymerization of the dye molecules by the tissue chromatropes

true metachromatic dyes
- toludine blue
- methylene blue

34
Q

alcian blue

A

stains acid mucin
at pH2.5 bith sulfated & carboxylated mucin is stained
at pH 1.0 only sulfated mucin stained

35
Q

Alcian blue -PAS allows us to be able to diagnose

A

Barretts esphagus

36
Q

alkaline congo red stains

A

amyloids
- builds up in conditons such as amyloidosis and eventually so much that it overtakes normal function of tissue and patient dies

37
Q

how to confirm that we see amyloid and not stain deposit

A

birefringent ( set up scope for polarization )

38
Q

what is used to remove unreduced silver in silver stains

A

hypo
sodium thiosulfate

39
Q

why do we reduce silver stains & example

A

reduces residual silver deposited at reaction site

ex. formaldehyde or hydroquinine

40
Q

what causes over staining in silver

A

if toning is carried out too long the backgroun dof the stained slide mat be violet to red color

41
Q

in masson trichchrome what do we post mordant in

A

bouins - to make more brilliant

42
Q

masson trichrome colors/ size

A

RBC : yellow
muscles: red
collagen : blue/green

43
Q

what does heteropolyacid do

A

 Phosphomolybdic acid (PMA) and/or

 Phosphotungstic acid (PTA)

“Colorless” anionic dyes

binds to tissue commponents
Collagen binds lots of PMA / PTA,

Heterpolyacid fits through collagen pores
and replaces the small anionic dye so collagen appears unstained

Large molecule dye bonds with the
heterpolyacid and stains collagen

44
Q

what is GMS used to stain

A

fungus* gold standard fungus stain

can also be used to stain urates

45
Q

where is fouchets reagent

A

bile stain

made up of ferris chloride and trichloracetic acid

46
Q

components of fouchets reagent

A

ferric chloride in trichloralacetic acid

47
Q

exogenous and endogenous pigments

A

endogenous

exogenous

48
Q

lipofuchin & bile pigment how to differentiate

A

lipofuschin is a wear & tear pigmnent
negative with bile stain fouchets

bile may accumulate when there is an obstruction
positive with bile fouchets stain ( green )

49
Q

melanin
how is it removed
how to stain
reducer or not

A

is an argentaffin substance thst doesnt require an extrsneous reducer so it it schmorl positive

removed by bleaching
not removed by weak acids

can use fontana masson to stain

50
Q

pearls prussian blue vs turnball

A

pearls prussian blue
- for ferric iron
- uses potassium ferrocyanide

turnball
- for ferrous iron ( rare)
- used in schmorl reaction
- uses ferricyanide to produce ferrous ferricyanide

51
Q

what stain can we use for melain and how to make it more sepcific

A

fontana masson

to make more specific
- make 2 slides and bleach one using potassium promagonate or hydrogen peroxide

52
Q

what methods are used for apud cells that are arrgarophilic

A

Grimelius & Churukin

both stains use hydroquinone as reducer & counterstained with nuclear fast red

IHC is more accurate means of ID

53
Q

exogenous pigments

A

carbon

abestos - birefringent
- coated with iron so can stain with pearls prussian blue

54
Q

calcium staining

A

stains blue/ purple in H& E

found abnormally in necrotic tissue

stain with
- von kossa * gold standard/ classic method ( uses light reduction )
- alizarin red

55
Q

urates what fixative should be avoided

A

water & lithium fixatives should be avoided

only use alcohol or frozen section

birefringent

GMS used to stain

56
Q

copper is associated with what disease and methods used

A

wilsons disease

rhodanine: more sensitive. less specific
rubeanic acid : less sensitive, more specific

57
Q

IHC least sensitive to most sensitive

A

direct (used for kidney or skin biopsies- use frozen sections )

indirect ( add patient serum to know antigen; looking for antinuclear antibodies )
- 2 step & 3 step

PAP( secondary & tertiary )

ABC & LSAB ( most specific )

58
Q

fixation of gynecological specimens

A

fixed immediately
ethanol
methanol can be used causes less shrinkage

dont use formalin

59
Q

nongyneological specimens

A

dont fix- bring to lab immeditaely

excess blood can obscure details

acetic acid in carnoys an clarkes lysis RBC- making it easier to ibserve nuclear detail

60
Q

liwuid based cytology two system

A

both produce high qulity monolayer slides
1. thin prep- uses filtration method
2. sure path liquid based pap test - monolayer preparations by sedimentation

61
Q

heparin

A

added to some body fluids to prevent clotting

62
Q

nickle method

A

breast/ nipple discharge

63
Q

CSf fluid

A

can deteriorate rapidly, if acant bring to lab immediately
use prefixative golding solution
- dilute specimen with an amount of alcoholic saline or saccomanno fluid equal to the volum eof the fluid

64
Q

urine for cytology

A

first morning not reccomennded

65
Q

pap stain 5 dyes 3 solutions

A

know these !

  1. hematoxylin ( nuclear stain)
  2. ORange G-6 first counterstain ( keratinized cells )
  3. EA36 ( polychrome cytoplasmic stain)
    - light green
    - eosin Y
    - Bismark brown
66
Q

cytology stain color results

A

chromatin - blue

keratin- orange

superficial squamous cells- variable shades of pink
nucleoli,cilia, RBCs - cells variable shades of pink

all metabolic cell cytoplasm cells- varibale shades of

Histology2 (14 decks)

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