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final review Flashcards

(66 cards)

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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
dark staining on edge
laser or electrocautery cannot fix burned tissue
26
poor contrast nuclei vs cytoplasm
poor nuclear staining excessive cytoplasmic staining
27
good technique for H&E
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
how to make & store schiff reagent
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
another name for PAS
McManus method
30
how to make PAS more specific for glycogen what fixative is preferred for glycogen
amylase or diastase = more speciific alcohol prevents glycogen streaming
31
whats a good control for glycogen
- liver - endo/ecto cervix
32
whats a good control for basement membrane
kidney
33
metachromatic stains
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
alcian blue
stains acid mucin at pH2.5 bith sulfated & carboxylated mucin is stained at pH 1.0 only sulfated mucin stained
35
Alcian blue -PAS allows us to be able to diagnose
Barretts esphagus
36
alkaline congo red stains
amyloids - builds up in conditons such as amyloidosis and eventually so much that it overtakes normal function of tissue and patient dies
37
how to confirm that we see amyloid and not stain deposit
birefringent ( set up scope for polarization )
38
what is used to remove unreduced silver in silver stains
hypo sodium thiosulfate
39
why do we reduce silver stains & example
reduces residual silver deposited at reaction site ex. formaldehyde or hydroquinine
40
what causes over staining in silver
if toning is carried out too long the backgroun dof the stained slide mat be violet to red color
41
in masson trichchrome what do we post mordant in
bouins - to make more brilliant
42
masson trichrome colors/ size
RBC : yellow muscles: red collagen : blue/green
43
what does heteropolyacid do
 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
what is GMS used to stain
fungus* gold standard fungus stain can also be used to stain urates
45
where is fouchets reagent
bile stain made up of ferris chloride and trichloracetic acid
46
components of fouchets reagent
ferric chloride in trichloralacetic acid
47
exogenous and endogenous pigments
endogenous exogenous
48
lipofuchin & bile pigment how to differentiate
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
melanin how is it removed how to stain reducer or not
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
pearls prussian blue vs turnball
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
what stain can we use for melain and how to make it more sepcific
fontana masson to make more specific - make 2 slides and bleach one using potassium promagonate or hydrogen peroxide
52
what methods are used for apud cells that are arrgarophilic
Grimelius & Churukin both stains use hydroquinone as reducer & counterstained with nuclear fast red IHC is more accurate means of ID
53
exogenous pigments
carbon abestos - birefringent - coated with iron so can stain with pearls prussian blue
54
calcium staining
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
urates what fixative should be avoided
water & lithium fixatives should be avoided only use alcohol or frozen section birefringent GMS used to stain
56
copper is associated with what disease and methods used
wilsons disease rhodanine: more sensitive. less specific rubeanic acid : less sensitive, more specific
57
IHC least sensitive to most sensitive
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
fixation of gynecological specimens
fixed immediately ethanol methanol can be used causes less shrinkage dont use formalin
59
nongyneological specimens
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
liwuid based cytology two system
both produce high qulity monolayer slides 1. thin prep- uses filtration method 2. sure path liquid based pap test - monolayer preparations by sedimentation
61
heparin
added to some body fluids to prevent clotting
62
nickle method
breast/ nipple discharge
63
CSf fluid
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
urine for cytology
first morning not reccomennded
65
pap stain 5 dyes 3 solutions
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
cytology stain color results
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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