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Histology2 > review for clinical exam > Flashcards

review for clinical exam Flashcards

1
Q

what stains are iron mordanted and what are they used for

A

verhoeffs
weigerts
celestine blue

used for acidic stains ( like trichrome)

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2
Q

what is the only suitable substitute for H& E

A

Celestine blue

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3
Q

bluing agent for H& E, and pH

A

scotts tap wate substitite pH 8

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4
Q

most important step before eosin & why

A

wash well bc bluing agent is pH 8 and eosin is acidic

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5
Q

what is eosins pH and its importance

A

pH 4.6-5 required for good staining with 3 different shades

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6
Q

differentiator for H&E ( if regressive )

A

1% HCl in 70% alcohol

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7
Q

regressive Hematoxylins

A

verhoeffs
delafield
Erlich

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8
Q

which stain cannot be hydrated of 70% and what does it stain

A

Aldehyde fuchsin ( for elastic stains ) is an alcoholic stain

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9
Q

what tissue doesnt need hydration

A

frozen sections bc it has no paraffin

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10
Q

what tissue doesnt need dehydration

A

metachromatic, & fat stains or anything else using an aqueous mounting media

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11
Q

metachromatic stains & what they are used to stain

A

methylene blue & toludene blue
stains mast cells, mucins

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12
Q

3 types of differentiators

A

excess mordant
oxidizer
weak alkali or acid

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13
Q

what is the oxidizer for H & E

A

sodium iodate

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14
Q

what type of hematoxylins dont require filtering & why

A

mayers ( chloral hydrate )
gills ( ethylene glycol)

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15
Q

which hematoxylin has a longer shelf life

A

aluminum mordanted (longer )
iron mordanted ( shorter)

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16
Q

what cant be used with eosin

A

isopropanol ( wont differentiate bc insoluble )

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17
Q

ways to test hematoxylin oxidiation and what it looks like over oxidized

A

filter paper ( maroon with purple edges)
few drops in water = blue/black
smells like wine

overoxidized = brown

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18
Q

colors of H&E

A

nuclei = blue/ black
calcium = blue/ black
everything else = shades of red
fungus = faintly stained

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19
Q

3 types of mordants

A

iron
aluminum ( most common )
tungsten ( special stains; muscle or CNS)

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20
Q

what does feulgen reaction stain and what color

A

DNA stained magenta

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21
Q

what fixative cant be used with feulgen

A

bouins

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22
Q

principle of feulgen reaction

A

hydrolyses of DNA with HCl = aldehyde group
aldehyde reacts with schiff reagent to color DNA

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23
Q

what is the control tissue for methyl- green pyronin Y

A

must contain plasma cells bc pyronin y is a plasma stain

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24
Q

principle of Methy green - pyronin Y stain and colors

A

differentiates DNA & RNA
DNA is more polymerized stains green ( methyl green)
RNA is less polymerized stains red ( pyronin Y )

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25
Q

may grunwald fixatives

A

zenker or B5

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26
Q

MAy grunwald is used to stain & control

A

bone marrow or hematopoetic cells
control= spleen

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27
Q

refractuve index of tissue and which type of mounting media matches it

A

~1.54
synthetic media = 1.51 - 1.55

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28
Q

PAS ( McManus ) oxidizer and what is produced

A

periodic acid oxidizes an aldehyde group

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29
Q

how is the schiff reagent made

A

parosaniline ( basic fuchsin) is treated with sulfuric acid which breaks quinoid ring = colorless ( leuco dye)

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30
Q

what stains use schiff reagent

A

PAS
feulgen
chromic acid schiff
Gridley
bielschowsky

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31
Q

how to make PAS more specific for glycogen

A

diastase or amylase

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32
Q

how to test schiffs regent

A

should be colorless
test with formaldehyde

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33
Q

what does mayers mucicarmine stain

A

epithelail mucins & cryptococcus neoformans

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34
Q

best control for glycogen

A

cervix or liver

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35
Q

what fixative cant be used with schiffs reagent

A

gluteraldehyde - causes false pos in PAS

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36
Q

what is the best fixative for glycogen

A

alcohol to prevent glycogen streaming

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37
Q

what color is PAS pos

A

magenta

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38
Q

what does bests carmine stain

A

glycogen

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39
Q

what does alcian blue stain

A

Acid mucins

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40
Q

pH uses of alcian blue

A

pH 2.5 stains sulfated & carboxylated mucins
pH 1 stains sulfated mucins

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41
Q

how to differentiate epithelial and connective tissue mucins

A

hylauronidase digests connective tissue mucins but leaves epithelial mucins

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42
Q

what does PAS Alcian blue differentiate

A

acid mucins ( alcian blue )
& neutral mucins ( PAS)

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43
Q

what disease does PAS & ALcian blue rule out

A

Barrettes esophagus

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44
Q

what does pearls prussian blue stain and what kind of reaction is it

A

stains iron blue
histochemical reaction

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45
Q

principle of pearls prussian blue

A

hemorsiderrin & HCl yields ferric chloride which reacts with potassium ferrocyanide = ferric ferrocyanide

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46
Q

when will you see amyloid build up

A

in certain pathological conditions gradually replacing cellular elements and leads to death

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47
Q

what are the most important forces in dye binding with amyloids

A

hydrophobic & van der walls

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48
Q

what was the original staining of amyloids with that is not specific

A

iodine

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49
Q

what is the most diffinitive way to stain amyloid

A

alkaline congo red

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50
Q

at is the key to diagnosis of amyloids

A

birefringence ; polarization makes more specific

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51
Q

section size for congo red stain

A

8-10 um

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52
Q

what is a good screen for amyloids

A

crystal violet ; may be used for frozen sections

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53
Q

staining method for crystal violet

A

polychromasia ; not well understood

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54
Q

what is the fluorescent dye for amyloids

A

thioflavine T

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55
Q

how big are sections for crystal violet stain

A

10-12 um

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56
Q

what are the types of collagen

A

major collagen
hyaline & elastin
recticular fibers
basement membrane

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57
Q

what is trichrome stain used to distinguish

A

collagen from muscle

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58
Q

what does increased collagen indicate

A

cirrhosis of the liver or renal disease

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59
Q

what is the pH of trichrome staining

A

1.5-3

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60
Q

what type of hematixylin should be used

A

iron hematoxyling ( verhoeffs or weigerts )

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61
Q

fixative for trichrome & how to make staining more brilliant

A

fixative if often formaldehyde ( the longer in formalin the less binding sites for stain )
mordant with picric acid ( bouins ) or mercuric chloride to inhance stain

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62
Q

what is the purpose of PTA & PMA

A

phosphotungstic acid & phosphomolybdic acid are colorless and cause smaller dyes to leave tissue ( collagen) to make room for bigger dyes, bigger dyes will bind to these acids

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63
Q

masson trichrome stain ( 3 sizes & colors) with examples of tissue

A

RBC ( small) yellow
muscle( med) red
collagen/ mucins ( large) blue

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64
Q

example of dense regular collagen & loose irregular collagen

A

dense regular = tendon
loose irregular = gallbladder

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65
Q

purpose of
collagen, reticular fiber, elastin,

A

collagen = strength
reticulin = supportive mesh
elastin = flexibility

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66
Q

if iron hematoxylin used what color is nucleus

A

black

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67
Q

how to stain basement membrane

A

carb technique or silver stains
PAS
JMS
GMS

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68
Q

what is the difference between bone and cartilage

A

cartilage lacunae are much larger

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69
Q

when is increased fibrin seen

A

tissue damage

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70
Q

what is the pH of reticulin stains

A

9

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71
Q

gomoris one step trichrome is mostly used for what &
colors

A

mostly used for muscle biopsies
colors identical to trichrome except all in one stain not 3 separate stains

72
Q

Van gieson stain contains why dyes

A

a trichrome stain containing picric acid & acid fuchsin

73
Q

colors for van gieson

A

cytoplasm & muscle = yellow
collagen = red
nuclei = black

74
Q

what does martius scarlette blue stain

A

trichrome stain for fibrin

75
Q

colors for MSB

A

martius yellow ( small )
nuclei = celestine blue or iron hematoxylin ( black )
crystal ponceau ( med dye) RED - FIBRIN
phosphotungstic acid
analine blue ( large )

76
Q

what is the oldest method for staining elastin

A

orcien

77
Q

what is the most widely used method for staining elastic

A

verhoeffs

78
Q

what does oxidation produce in specific elastin stains

A

sulphonic acid derivatives = basophilic

79
Q

what does aldehyde fuchsin stain

A

elastin

80
Q

what is the oxidizer for verhoeffs

A

iodine = oxidizer & mordant

81
Q

what kind of stain is verhoeffs & what is its differentiator

A

regressive
ferric chloride = differentiator & mordant

82
Q

what is used to remove iodine in verhoeffs

A

sodium thio sulfate

83
Q

what stain is alcholic & shouldnt be hydrated beyond 70%

A

aldehyde fuchsin

84
Q

what is in aldehyde fuchsin stain

A

alcoholic parosaniline, HCl & paraldehyde

85
Q

what is the oxidizer for aldehyde fuchsin

A

permanganate

86
Q

movat pentachrome stains & tissue stained

A

nuclei & elastin = black ( verhoeffs)
collagen = yellow ( saffron )
ground substance & mucin = blue ( alcian blue)
fibrin & muscle = red ( acid fuchsin)
[hosphotungstic acid = colorless

87
Q

what are silver stains known as

A

metallic impregnation; no dye metallic ions deposited

88
Q

main metal in silver stains

A

silver

89
Q

uses of metallic reduction

A

reticular fibers
bacteria ( spirochetes)
fungi
neuropathology ( aldehydes, calcium)

90
Q

what are 4 important things to remember for silver stains

A
  1. must be neutralized before dicarding or may become explosive
  2. never used metal forcepts
  3. use chemically clean glassware
  4. sections tend to float off slide due to alkalinity ( use histogrip)
91
Q

argyrophilic vs argentaffin

A

“phil” needs an extraneous reducer
“finn” doesn’ t

92
Q

purpose of
oxidizer, sensitizer, bleach, toner, fixer, reducer

A

oxidizer= produces aldehydes ( potassium permanganate
bleach =removes purple from oxidizer ( oxalic acid)
sensitizer = enhances silver deposits
toner = produces contrast (gold)
fixer= removes unreduced silver ( sodium thio)
reducer= used in argentafin methods to turn silver black ( formaldehyde or hydroquinone)

93
Q

what does gomoris or gordon & sweets stain & what kind of stain is it

A

reticulin
agarophilic silver stain

94
Q

ingredients of gomoris or gordon & sweets

A

oxidizer = potassium permanganate
bleach = oxalic acid
silver solution
sensitizer = ferric ammonium sulfate
toner= gold
reducer= formaldehyde
potassium metabisulfate ( gormoris)
fixer= sodium thio

95
Q

what does mallory PTAH stain

A

muscle striations, fibrin, glial fibers, brain tissue

96
Q

what mordant, fixative & oxidizer is used in PTAH

A

hematoxylin with tungsten mordant
zenker fixative
permenganate oxidizer

97
Q

what does JMS stain & its oxidizer

A

basement membrane & reticulin fibers
oxidizer is periodic acid

98
Q

how do lipid stains work and what are they called

A

lysochrome stains
the dye is more soluble in lipid than the solvent

99
Q

what solvents are used in lipid stains

A

isopropanol or propylene glycol ( not water )

100
Q

2 non common lipid stains

A

sudan black & oil red O

101
Q

lipid stain if parrafin section used

A

osmium tetraoxide

102
Q

what does osmium tetraoxide do to lipids

A

preserves fat & turns them black

103
Q

what is nissl substance & what is it made up of

A

made of rough endoplastmic redticulum & RNA, in neuron cytoplasm

104
Q

chromatolysis

A

during injury the nissl substance starts to dissapear around the nuclei first

105
Q

what do you stain nissl substance with

A

cresyl echt violet

106
Q

what is used to stain myelin

A

luxol fast blue or iron hematocylin ( most is lost through processing )

107
Q

what kind of method is Bodian & what are the ingredients

A

agyrophilic
protargoltm ( contains copper)
gold
oxalic acid
hydroquinone & formaldehyde
sodium thio

108
Q

what kind of method is holmes & section size

A

agarlophilic - reduce with hydroquinone & sodium sulfite
10-15 um

109
Q

what does bielschowsky stain

A

neurofibilary tangles
if used w/ PAS - will stain amyloid in senile plaques

110
Q

ingredients in bielschowsky

A

silver
gold
hypo
periodic acid
schiffs

111
Q

what does Gallyas stain

A

tau protiens

112
Q

ingredients of gallyas stain

A

periodic acid
siliver iodide
acetic acid
gold
thio
nuclear fast red

113
Q

what does gallyas not detect

A

amyloids

114
Q

what color does PTAH stain neurons

A

neurons = salmon
evrything else = blue

115
Q

what is in Holzer method

A

crystal violet ( glial fibers)
cholorform ( decolorizer)
= toxic soup

116
Q

what does Cajal stain

A

astrocytes using metal impregnation

117
Q

what are astrocytes

A

normal but lots means brain is trying to repair

118
Q

what is a relative of alcian blue

A

luxol fast blue

119
Q

what does weil stain

A

myelin

120
Q

weil ingredients

A

iron hematoxylin
ferric ammonium sulfate ( mordant)
sodium borate potassium ferrocyanide ( oxidizer)

121
Q

what is important with brain slides

A

tend to come off ( use albumin)
dry overnight at 37 degrees
sections 6-8 um

122
Q

things used to ID alzehimers

A

tau proteins
senile plaques ( amyloid)
neurofibrillary tangles

123
Q

what are the disinfectants for CJD

A

phenol & chlorine

124
Q

what is the fixative for CJD

A

formalin & formic acid *dont mix chlorine & formalin ( toxic)

125
Q

where is the danger of infection in histology

A

autopsy
grossing
frozen section s

126
Q

what fixative cant be used for AFB

A

carnoys ( alcohol removes acid fastness)

127
Q

what techniques are used for AFB

A

lower surface tension forcing stain into capsule
- phenol
- heat
- tergitol

128
Q

what does kinyoun stain

A

diagnostic for AFB

129
Q

what is in kinyoun stain

A

basic fuchsin, melted phenol crystals
- usually heated
differentiate in acid alcohol

130
Q

whats makes ziehl neilson different from kinyoun

A

same as kinyoun but much less concentrated
microwave method

131
Q

what stain is used for leprosy

A

fites stain

132
Q

what is special about m.leprae

A

capsule lost in clearing; preserve with peanut oil

133
Q

what are fluorescent dyes for AFB

A

Auramine & RHodamine( good for screening )
- good for few organisms

134
Q

procedure for auramine & rhodamine

A

heat forces dye in
differentiate in acid alcohol
stain in eriochome Black T or quench in potsssium permenganate ( quenching background fluorescence)

135
Q

colors in kinyoun stain

A

AFB = red
background- metheylene blue

136
Q

what does brown & hopps stain

A

RED= gram neg bacteria & ricketsia

137
Q

what does brown & brenn stain

A

BLUE = gram pos bacteria

138
Q

what happens if you allow sections to dry

A

insoluble compound formed

139
Q

what is the differentiatir in brown & hopps

A

gallego

140
Q

what does giemsa stain

A

H.pylori, ricketsia

141
Q

what are the 4 fungus stains from least specific to most

A

hotchkiss- mcmanus (PAS)
chromic acid schiff
Gridley
GMS

142
Q

how to clean up background in fungi PAS Stain

A

diastase gets rid of glycogen

143
Q

what is the oxidizer in CAS and what does it do

A

chromic acid eliminates aldehydes from all carbohydrates except those of the highest concentration
lowest ( mucin, glycogen)
highest ( fungal cell wall)

144
Q

what is in the gridley stain

A

schiff regaent
chromic acid
aldehyde fuchsin ( makes stronger rxn)

145
Q

what is the pH buffer in GMS stain

A

sodium tetraborax

146
Q

what color is fungi in GMS

A

black bc silver stain

147
Q

what causes background staining in GMS

A

heating above 62 degrees or staining too long

148
Q

what is the oxidizer, bleach & toner in GMS

A

chromic acid oxidizer
sodium bisulfate bleach
gold toner ( changes from brown to black)

149
Q

what is the heating step in GMS

A

heat at 56 until paper bag brown ( yellow- brown -black) if red over toned

150
Q

what does mayers mucicarmine stain

A

wall of fungus & capsule of cryptococcus neoformans

151
Q

what kind of stain is warthin starry & what does it stain

A

agarophilic stain for spirochettes

152
Q

what does warthin starry involve & what color does it stain

A

lots of prep work & pre warming
stains spirochettes black
background yellow

153
Q

what water is used for warthin starry

A

acidulated water - triple distilled

154
Q

what kind of reaction is prussian blue & what does it stain

A

a hsitochemical reaction that stains ferric iron

155
Q

principle of prussian blue stain

A

potassium ferrocyanide + HCl =ferric ferrocyanide
which detects hemosiderin ( iron)

156
Q

control for prussian blue

A

limited iron in control to avoid background stain

157
Q

what is the turnball reaction used in

A

the shmorl technique

158
Q

what does the turnball reaction stain

A

ferrous iron
using ferrous ferricyanide

159
Q

what are the precautions for prussina blue/ turnball reaction

A

chemically clean glassware
non metal forecepts

160
Q

what does the shmorl technique stain

A

melanin
argentafin granules
formalin pigment

161
Q

what kind of reaction is fontanna masson & what does it stain

A

silver argentaffin technique for melanin & APUD cells

162
Q

what can be done to make melanin stain more specific

A

in fontanna masson make 2 slides, bleach one of the slides

163
Q

what removes melanin & calcium

A

melanin = bleach
calcium = sulfuric acid

164
Q

what are 2 silver methods for APUD cells that are agyrophilic

A

GRimelius & Churukian
hydroquinone reducer

165
Q

stain & fixative for urates

A

GMS - same as fungi but stain longer
alcohol fixative
frozen sections & polarizer can be used instead

166
Q

what can urates cause

A

gout

167
Q

what are the 3 pigments produced by stains

A

formic acid
mercuric
chromate

168
Q

how do the 3 pigments from stains occur

A

formic acid= ph<6 and blood
mercuric= unavoidable
chromate = not rinsing after fixation

169
Q

how to remove pigments

A

formic acid = acoholic picric acid or alcoholic alkali
mercuric= iodine & sodium thio
chromate = cant ( some say acid alcohol)

170
Q

what is the most common pigment in tisse

A

carbon

171
Q

what are abestos bodies

A

dumbell shaped, coated in iron ( stains with prussian blue )
birefringent

172
Q

how to tell the difference between bile and lipofuchsin & COLORS

A

bile stain
bile = green
bakground = yellow

173
Q

what is in bile stain & what it does

A

fouchets reagent ( ferric chloride & tetrchloracetic acid ) oxidizes bilirubin to biliverdin

174
Q

bile vs lipofuchsin

A

bile = dangerous, builds up with obstruction
lipofuchsin = wear & tear pigment

175
Q

what does von kossa stain & what type of stain is it, what color

A

silver stain for calcium

176
Q

what does alzarin red stain & pH ( how to maintain pH )

A

pH 4.1-4.3 makes it specific for calcium ( use ammonium hydroxide to get certain pH

177
Q

what is copper stain for & 2 methods

A

wilsons disease
rhodanine= less specific & more sensistive
rubeanic = more specific & less sensitive

Histology2 (14 decks)

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