review for clinical exam Flashcards
(177 cards)
1
Q
what stains are iron mordanted and what are they used for
A
verhoeffs
weigerts
celestine blue
used for acidic stains ( like trichrome)
2
Q
what is the only suitable substitute for H& E
A
Celestine blue
3
Q
bluing agent for H& E, and pH
A
scotts tap wate substitite pH 8
4
Q
most important step before eosin & why
A
wash well bc bluing agent is pH 8 and eosin is acidic
5
Q
what is eosins pH and its importance
A
pH 4.6-5 required for good staining with 3 different shades
6
Q
differentiator for H&E ( if regressive )
A
1% HCl in 70% alcohol
7
Q
regressive Hematoxylins
A
verhoeffs
delafield
Erlich
8
Q
which stain cannot be hydrated of 70% and what does it stain
A
Aldehyde fuchsin ( for elastic stains ) is an alcoholic stain
9
Q
what tissue doesnt need hydration
A
frozen sections bc it has no paraffin
10
Q
what tissue doesnt need dehydration
A
metachromatic, & fat stains or anything else using an aqueous mounting media
11
Q
metachromatic stains & what they are used to stain
A
methylene blue & toludene blue
stains mast cells, mucins
12
Q
3 types of differentiators
A
excess mordant
oxidizer
weak alkali or acid
13
Q
what is the oxidizer for H & E
A
sodium iodate
14
Q
what type of hematoxylins dont require filtering & why
A
mayers ( chloral hydrate )
gills ( ethylene glycol)
15
Q
which hematoxylin has a longer shelf life
A
aluminum mordanted (longer )
iron mordanted ( shorter)
16
Q
what cant be used with eosin
A
isopropanol ( wont differentiate bc insoluble )
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
18
Q
colors of H&E
A
nuclei = blue/ black
calcium = blue/ black
everything else = shades of red
fungus = faintly stained
19
Q
3 types of mordants
A
iron
aluminum ( most common )
tungsten ( special stains; muscle or CNS)
20
Q
what does feulgen reaction stain and what color
A
DNA stained magenta
21
Q
what fixative cant be used with feulgen
A
bouins
22
Q
principle of feulgen reaction
A
hydrolyses of DNA with HCl = aldehyde group
aldehyde reacts with schiff reagent to color DNA
23
Q
what is the control tissue for methyl- green pyronin Y
A
must contain plasma cells bc pyronin y is a plasma stain
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 )
25
may grunwald fixatives
zenker or B5
26
MAy grunwald is used to stain & control
bone marrow or hematopoetic cells
control= spleen
27
refractuve index of tissue and which type of mounting media matches it
~1.54
synthetic media = 1.51 - 1.55
28
PAS ( McManus ) oxidizer and what is produced
periodic acid oxidizes an aldehyde group
29
how is the schiff reagent made
parosaniline ( basic fuchsin) is treated with sulfuric acid which breaks quinoid ring = colorless ( leuco dye)
30
what stains use schiff reagent
PAS
feulgen
chromic acid schiff
Gridley
bielschowsky
31
how to make PAS more specific for glycogen
diastase or amylase
32
how to test schiffs regent
should be colorless
test with formaldehyde
33
what does mayers mucicarmine stain
epithelail mucins & cryptococcus neoformans
34
best control for glycogen
cervix or liver
35
what fixative cant be used with schiffs reagent
gluteraldehyde - causes false pos in PAS
36
what is the best fixative for glycogen
alcohol to prevent glycogen streaming
37
what color is PAS pos
magenta
38
what does bests carmine stain
glycogen
39
what does alcian blue stain
Acid mucins
40
pH uses of alcian blue
pH 2.5 stains sulfated & carboxylated mucins
pH 1 stains sulfated mucins
41
how to differentiate epithelial and connective tissue mucins
hylauronidase digests connective tissue mucins but leaves epithelial mucins
42
what does PAS Alcian blue differentiate
acid mucins ( alcian blue )
& neutral mucins ( PAS)
43
what disease does PAS & ALcian blue rule out
Barrettes esophagus
44
what does pearls prussian blue stain and what kind of reaction is it
stains iron blue
histochemical reaction
45
principle of pearls prussian blue
hemorsiderrin & HCl yields ferric chloride which reacts with potassium ferrocyanide = ferric ferrocyanide
46
when will you see amyloid build up
in certain pathological conditions gradually replacing cellular elements and leads to death
47
what are the most important forces in dye binding with amyloids
hydrophobic & van der walls
48
what was the original staining of amyloids with that is not specific
iodine
49
what is the most diffinitive way to stain amyloid
alkaline congo red
50
at is the key to diagnosis of amyloids
birefringence ; polarization makes more specific
51
section size for congo red stain
8-10 um
52
what is a good screen for amyloids
crystal violet ; may be used for frozen sections
53
staining method for crystal violet
polychromasia ; not well understood
54
what is the fluorescent dye for amyloids
thioflavine T
55
how big are sections for crystal violet stain
10-12 um
56
what are the types of collagen
major collagen
hyaline & elastin
recticular fibers
basement membrane
57
what is trichrome stain used to distinguish
collagen from muscle
58
what does increased collagen indicate
cirrhosis of the liver or renal disease
59
what is the pH of trichrome staining
1.5-3
60
what type of hematixylin should be used
iron hematoxyling ( verhoeffs or weigerts )
61
fixative for trichrome & how to make staining more brilliant
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
62
what is the purpose of PTA & PMA
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
63
masson trichrome stain ( 3 sizes & colors) with examples of tissue
RBC ( small) yellow
muscle( med) red
collagen/ mucins ( large) blue
64
example of dense regular collagen & loose irregular collagen
dense regular = tendon
loose irregular = gallbladder
65
purpose of
collagen, reticular fiber, elastin,
collagen = strength
reticulin = supportive mesh
elastin = flexibility
66
if iron hematoxylin used what color is nucleus
black
67
how to stain basement membrane
carb technique or silver stains
PAS
JMS
GMS
68
what is the difference between bone and cartilage
cartilage lacunae are much larger
69
when is increased fibrin seen
tissue damage
70
what is the pH of reticulin stains
9
71
gomoris one step trichrome is mostly used for what &
colors
mostly used for muscle biopsies
colors identical to trichrome except all in one stain not 3 separate stains
72
Van gieson stain contains why dyes
a trichrome stain containing picric acid & acid fuchsin
73
colors for van gieson
cytoplasm & muscle = yellow
collagen = red
nuclei = black
74
what does martius scarlette blue stain
trichrome stain for fibrin
75
colors for MSB
martius yellow ( small )
nuclei = celestine blue or iron hematoxylin ( black )
crystal ponceau ( med dye) RED - FIBRIN
phosphotungstic acid
analine blue ( large )
76
what is the oldest method for staining elastin
orcien
77
what is the most widely used method for staining elastic
verhoeffs
78
what does oxidation produce in specific elastin stains
sulphonic acid derivatives = basophilic
79
what does aldehyde fuchsin stain
elastin
80
what is the oxidizer for verhoeffs
iodine = oxidizer & mordant
81
what kind of stain is verhoeffs & what is its differentiator
regressive
ferric chloride = differentiator & mordant
82
what is used to remove iodine in verhoeffs
sodium thio sulfate
83
what stain is alcholic & shouldnt be hydrated beyond 70%
aldehyde fuchsin
84
what is in aldehyde fuchsin stain
alcoholic parosaniline, HCl & paraldehyde
85
what is the oxidizer for aldehyde fuchsin
permanganate
86
movat pentachrome stains & tissue stained
nuclei & elastin = black ( verhoeffs)
collagen = yellow ( saffron )
ground substance & mucin = blue ( alcian blue)
fibrin & muscle = red ( acid fuchsin)
[hosphotungstic acid = colorless
87
what are silver stains known as
metallic impregnation; no dye metallic ions deposited
88
main metal in silver stains
silver
89
uses of metallic reduction
reticular fibers
bacteria ( spirochetes)
fungi
neuropathology ( aldehydes, calcium)
90
what are 4 important things to remember for silver stains
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
argyrophilic vs argentaffin
"phil" needs an extraneous reducer
"finn" doesn' t
92
purpose of
oxidizer, sensitizer, bleach, toner, fixer, reducer
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
what does gomoris or gordon & sweets stain & what kind of stain is it
reticulin
agarophilic silver stain
94
ingredients of gomoris or gordon & sweets
oxidizer = potassium permanganate
bleach = oxalic acid
silver solution
sensitizer = ferric ammonium sulfate
toner= gold
reducer= formaldehyde
potassium metabisulfate ( gormoris)
fixer= sodium thio
95
what does mallory PTAH stain
muscle striations, fibrin, glial fibers, brain tissue
96
what mordant, fixative & oxidizer is used in PTAH
hematoxylin with tungsten mordant
zenker fixative
permenganate oxidizer
97
what does JMS stain & its oxidizer
basement membrane & reticulin fibers
oxidizer is periodic acid
98
how do lipid stains work and what are they called
lysochrome stains
the dye is more soluble in lipid than the solvent
99
what solvents are used in lipid stains
isopropanol or propylene glycol ( not water )
100
2 non common lipid stains
sudan black & oil red O
101
lipid stain if parrafin section used
osmium tetraoxide
102
what does osmium tetraoxide do to lipids
preserves fat & turns them black
103
what is nissl substance & what is it made up of
made of rough endoplastmic redticulum & RNA, in neuron cytoplasm
104
chromatolysis
during injury the nissl substance starts to dissapear around the nuclei first
105
what do you stain nissl substance with
cresyl echt violet
106
what is used to stain myelin
luxol fast blue or iron hematocylin ( most is lost through processing )
107
what kind of method is Bodian & what are the ingredients
agyrophilic
protargoltm ( contains copper)
gold
oxalic acid
hydroquinone & formaldehyde
sodium thio
108
what kind of method is holmes & section size
agarlophilic - reduce with hydroquinone & sodium sulfite
10-15 um
109
what does bielschowsky stain
neurofibilary tangles
if used w/ PAS - will stain amyloid in senile plaques
110
ingredients in bielschowsky
silver
gold
hypo
periodic acid
schiffs
111
what does Gallyas stain
tau protiens
112
ingredients of gallyas stain
periodic acid
siliver iodide
acetic acid
gold
thio
nuclear fast red
113
what does gallyas not detect
amyloids
114
what color does PTAH stain neurons
neurons = salmon
evrything else = blue
115
what is in Holzer method
crystal violet ( glial fibers)
cholorform ( decolorizer)
= toxic soup
116
what does Cajal stain
astrocytes using metal impregnation
117
what are astrocytes
normal but lots means brain is trying to repair
118
what is a relative of alcian blue
luxol fast blue
119
what does weil stain
myelin
120
weil ingredients
iron hematoxylin
ferric ammonium sulfate ( mordant)
sodium borate potassium ferrocyanide ( oxidizer)
121
what is important with brain slides
tend to come off ( use albumin)
dry overnight at 37 degrees
sections 6-8 um
122
things used to ID alzehimers
tau proteins
senile plaques ( amyloid)
neurofibrillary tangles
123
what are the disinfectants for CJD
phenol & chlorine
124
what is the fixative for CJD
formalin & formic acid *dont mix chlorine & formalin ( toxic)
125
where is the danger of infection in histology
autopsy
grossing
frozen section s
126
what fixative cant be used for AFB
carnoys ( alcohol removes acid fastness)
127
what techniques are used for AFB
lower surface tension forcing stain into capsule
- phenol
- heat
- tergitol
128
what does kinyoun stain
diagnostic for AFB
129
what is in kinyoun stain
basic fuchsin, melted phenol crystals
- usually heated
differentiate in acid alcohol
130
whats makes ziehl neilson different from kinyoun
same as kinyoun but much less concentrated
microwave method
131
what stain is used for leprosy
fites stain
132
what is special about m.leprae
capsule lost in clearing; preserve with peanut oil
133
what are fluorescent dyes for AFB
Auramine & RHodamine( good for screening )
- good for few organisms
134
procedure for auramine & rhodamine
heat forces dye in
differentiate in acid alcohol
stain in eriochome Black T or quench in potsssium permenganate ( quenching background fluorescence)
135
colors in kinyoun stain
AFB = red
background- metheylene blue
136
what does brown & hopps stain
RED= gram neg bacteria & ricketsia
137
what does brown & brenn stain
BLUE = gram pos bacteria
138
what happens if you allow sections to dry
insoluble compound formed
139
what is the differentiatir in brown & hopps
gallego
140
what does giemsa stain
H.pylori, ricketsia
141
what are the 4 fungus stains from least specific to most
hotchkiss- mcmanus (PAS)
chromic acid schiff
Gridley
GMS
142
how to clean up background in fungi PAS Stain
diastase gets rid of glycogen
143
what is the oxidizer in CAS and what does it do
chromic acid eliminates aldehydes from all carbohydrates except those of the highest concentration
lowest ( mucin, glycogen)
highest ( fungal cell wall)
144
what is in the gridley stain
schiff regaent
chromic acid
aldehyde fuchsin ( makes stronger rxn)
145
what is the pH buffer in GMS stain
sodium tetraborax
146
what color is fungi in GMS
black bc silver stain
147
what causes background staining in GMS
heating above 62 degrees or staining too long
148
what is the oxidizer, bleach & toner in GMS
chromic acid oxidizer
sodium bisulfate bleach
gold toner ( changes from brown to black)
149
what is the heating step in GMS
heat at 56 until paper bag brown ( yellow- brown -black) if red over toned
150
what does mayers mucicarmine stain
wall of fungus & capsule of cryptococcus neoformans
151
what kind of stain is warthin starry & what does it stain
agarophilic stain for spirochettes
152
what does warthin starry involve & what color does it stain
lots of prep work & pre warming
stains spirochettes black
background yellow
153
what water is used for warthin starry
acidulated water - triple distilled
154
what kind of reaction is prussian blue & what does it stain
a hsitochemical reaction that stains ferric iron
155
principle of prussian blue stain
potassium ferrocyanide + HCl =ferric ferrocyanide
which detects hemosiderin ( iron)
156
control for prussian blue
limited iron in control to avoid background stain
157
what is the turnball reaction used in
the shmorl technique
158
what does the turnball reaction stain
ferrous iron
using ferrous ferricyanide
159
what are the precautions for prussina blue/ turnball reaction
chemically clean glassware
non metal forecepts
160
what does the shmorl technique stain
melanin
argentafin granules
formalin pigment
161
what kind of reaction is fontanna masson & what does it stain
silver argentaffin technique for melanin & APUD cells
162
what can be done to make melanin stain more specific
in fontanna masson make 2 slides, bleach one of the slides
163
what removes melanin & calcium
melanin = bleach
calcium = sulfuric acid
164
what are 2 silver methods for APUD cells that are agyrophilic
GRimelius & Churukian
hydroquinone reducer
165
stain & fixative for urates
GMS - same as fungi but stain longer
alcohol fixative
frozen sections & polarizer can be used instead
166
what can urates cause
gout
167
what are the 3 pigments produced by stains
formic acid
mercuric
chromate
168
how do the 3 pigments from stains occur
formic acid= ph<6 and blood
mercuric= unavoidable
chromate = not rinsing after fixation
169
how to remove pigments
formic acid = acoholic picric acid or alcoholic alkali
mercuric= iodine & sodium thio
chromate = cant ( some say acid alcohol)
170
what is the most common pigment in tisse
carbon
171
what are abestos bodies
dumbell shaped, coated in iron ( stains with prussian blue )
birefringent
172
how to tell the difference between bile and lipofuchsin & COLORS
bile stain
bile = green
bakground = yellow
173
what is in bile stain & what it does
fouchets reagent ( ferric chloride & tetrchloracetic acid ) oxidizes bilirubin to biliverdin
174
bile vs lipofuchsin
bile = dangerous, builds up with obstruction
lipofuchsin = wear & tear pigment
175
what does von kossa stain & what type of stain is it, what color
silver stain for calcium
176
what does alzarin red stain & pH ( how to maintain pH )
pH 4.1-4.3 makes it specific for calcium ( use ammonium hydroxide to get certain pH
177
what is copper stain for & 2 methods
wilsons disease
rhodanine= less specific & more sensistive
rubeanic = more specific & less sensitive
