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Histopathology Lec&Lab > MODULE 5 - Staining & Mounting > Flashcards

MODULE 5 - Staining & Mounting Flashcards

1
Q

Process of applying dyes on the section to see and study the architectural pattern of the tissue and physical characteristics of cells

A

staining

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2
Q

Tissue components are demonstrated by direct interaction with a dye or staining solution

A

Histological stain

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3
Q

used for chemical component/reaction

A

Histochemical stain

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4
Q

Active tissue component is colored

A

Histological stain

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5
Q

Examples of histological stains

A

Micro-anatomical stains
Bacterial stains
Specific tissue stains

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6
Q

Uses antibodies instead of stains

A

Immunohistochemical stain

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7
Q

Perl’s Prussian blue reaction is for

A

hemoglobin

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8
Q

Periodic acid Schiff is for

A

Carbohydrates

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9
Q

Examples of histochemical stains

A

Perl’s prussian blue

Periodic and schiff

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10
Q

Detection of phenotypic markers that are detected by antibodies

A

immunohistochemical stain

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11
Q

Examples of immunohistochemical staining

A

monoclonal

polyclonal

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12
Q

uses aqueous or alcoholic dye solutions to produce a color

A

Direct staining

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13
Q

Uses only one dye, usual color is the resulting color

A

direct staining

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14
Q

Example of direct staining

A

methylene blue

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15
Q

Uses mordant/accentuator

A

Indirect staining

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16
Q

Link/bridge between dye and tissue

A

mordant

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17
Q

Examples of mordant and solutions that contains them

A

Potassium alum- Meyer’s hematoxylin

Iron (Ferric ammonium chloride)- Weigert’s hematoxylin

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18
Q

What is accentuator

A

It heightens the color intensity and selectivity

Increases staining power

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19
Q

Examples of accentuator and solutions that contains them

A

KOH- Loeffler’s methylene blue

Phenol- carbol fuschin or carbol thionine

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20
Q

tissue elements re stained in a definite sequence
gradual application of dyes
No excess dye No decolorization

A

Progressive staining

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21
Q

Tissue is overstained

A

Regressive staining

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22
Q

Excess dye in regressive staining are removed by

A

Decolorization/differentiation

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23
Q

Example of regressive staining

A

Acid alcohol- can remove both acid and basic dyes

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24
Q

staining tissue with a color that is different from the stain itself

A

Metachromatic staining

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25
Q

Examples of metachromatic staining

A 
Methyl violet or crystal violet
Cresyl blue for reticulocytes
Safranin
Bismarck brown
Basic fuschin
Methylene blue
Thionine
Toluidine blue
Azure A,B,C
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26
Q

involves application of a different color to produce contrast background

A

Counterstaining

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27
Q

Example of counterstain and explain its use

A

Eosin- red acid dye for cytoplasmic stain

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28
Q

Forms of eosin

A

✓ Eosin Y- yellow
✓ Bluish- eosin b
✓ Ethyl Eosin- eosin s

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29
Q

tissue elements are demonstrated NOT by stains but by colorless solutions of metallic salts

A

Metallic impregnation

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30
Q

When applied to tissues, it is not absorbed and can only leave black deposits on surface

A

Metallic impregnation

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31
Q

Examples of metallic impregnation

A

Ammoniacal silver
Silver nitrate
Gold chloride

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32
Q

Demonstrates living cells, fresh and viable cells.

A

Vital staining

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33
Q

2 types of vital staining

A

Intravital

Supravital

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34
Q

Dye is injected to any part of the body

A

Intravital

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35
Q

Examples of intravital stains

A

Lithium
Carmine
India ink

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36
Q

stain is applied immediately to tissue after its removal from living body

A

Supravital

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37
Q

Best vital stain

A

Neutral red

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38
Q

Supravital stain for mitochondria

A

Janus green

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39
Q

Examples of supravital stain

A 
Neutral red- best  vital stain
Janus green- for mitochondria
Trypan blue
Nile blue
Toluidine blue
Thionine
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40
Q

Most common method utilized for microanatomical studies of tissues

A

Rountine H&E

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41
Q

H&E stain is what type of stain

A

Regressive staining

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42
Q

Steps in H&E staining

A
  1. Initial xylene bath- for further deparaffinization
  2. Descending grades of alcohol- for hydration
  3. Hematoxylin- primary dye, nuclear stain-basic dye
  4. Acid alcohol- decolorizer
  5. Ammonia water- bluing agent
  6. Eosin- counterstain, cytoplasmic stain-acid dye
  7. Ascending grades of alcohol- for dehydration
  8. Final xylene bath- for clearing prior to mounting
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43
Q

Enumerate H&E results

A

Nuclei- blue to blue black
Karyosome- deep blue
Cytoplasm- pale pink
RBCs, eosinophilic granules, keratin- bright orange red
Calcium and decalcified bones- purplish blue
Decalcified bone matrix, osteoid, collagen- pink
Muscle tissue- deep pink

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44
Q

Staining Methods for FROZEN SECTION

A

H&E
Thionine
Polychrome methylene blue
Alcohol pinacyanol method

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45
Q

– derived from plants and animals

A

natural dyes

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46
Q

Examples of natural dyes

A

Hematoxylin
Cochineal dyes
Orcein
Saffron

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47
Q

Hematoxylin is a stain

True or False

A

False

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48
Q

active coloring agent of hematoxylin

A

Hematane

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49
Q

Hematoxylin is formed from the oxidation of hematoxylin either by

A

ripening (Exposure to sunlight)

adding oxidizing agents

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50
Q

Oxidizing agents in hematoxylin

A 
Hydrogen peroxide
Mercuric peroxide
Sodium iodate
Sodium perborate 
Potassium permanganate
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51
Q

Hematoxylin is used in combination with

A

alum
iron
chromium
copper salts

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52
Q

ALUM HEMATOXYLIN uses _______ as mordant

A

potassium alum

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53
Q

Ripening agent of Ehrlich’s hematoxylin

A

Sodium iodate

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54
Q

Ripening agent of Harris hematoxylin

A

Mercuric oxide- for exfoliative cytology and sec chromosomes

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55
Q

Ripening agent of Cole’s hematoxylin

A

Alcoholic iodine

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56
Q

Ripening agent of Mayer’s hematoxylin

A

Sodium iodate
Contains citric acid and chloral hydrate as preservative
preferred for immunohistochem

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57
Q

Ripening agent of Delafield’s hematoxylin

A

Glycerol- to stabilize oxidation process

Smells like wine, color purple-red shade

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58
Q

Ripening agent of Gill’s hematoxylin

A

sodium iodate

used to stain mucin in goblet’s cell

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59
Q

Ripening agent of Carazzi’s hematoxylin

A

Potassium iodate

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60
Q

– in general for photomicrography

A

IRON HEMATOXYLIN

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61
Q

Weigert’s Hematoxylin is for?

and what is its component/mordant

A

Muscle fibers and CT

Ferric ammonium chloride- both mordant and ripening agent

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62
Q

Heidenhain’s Hematoxylin is for?

and what is its component/mordant

A

nucleus and cytoplasm (mitochondria)

Ferric ammonium sulfate- both mordant and ripening agent

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63
Q

COPPER HEMATOXYLIN is used for

A

spermatogenesis

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64
Q

TUNGSTEN HEMATOXYLIN contains

A

phosphotungstic acid

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65
Q

LEAD HEMATOXYLIN is for

A

demo of granules of endocrine cells of alimentary tract

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66
Q

MOLYBDENUM HEMATOXYLIN is for

A

demo of collagen, reticulin and argentaffin cells

67
Q

extracted from bug treated with alum to produce carmine

A

Cochineal dyes

68
Q

Cochineal dyes is treated with alum to produce

A

carmine

69
Q

Cochineal dyes with picric acid will produce

A

picro carmine- for neuropathological studies

70
Q

Cochineal dyes with aluminum chloride will produce

A

Bescarmine- glycogen

71
Q

– vegetable dye extracted from lichens and is used for ______

A

Orcein

staining elastic fibers

72
Q

vegetable dye extracted from lichens to be used as pH indicator because of poor staining property

A

Litmus

73
Q

Synthetic dyes also known as

A

Coar tal/aniline dyes

74
Q

2 components of Synthetic dyes

A
  1. Chromophore- coloring property

2. Auxochrome- dyeing propert, to retain the imparted color

75
Q

what is lysochrome dyes

A

also known as Oil soluble dyes, used for FATS

It has no auxochrome

76
Q

Most sensitive soluble dye

A

Sudan black B

77
Q

Sudan IV also known as and what is its use

A

Sharlach R

Neutral lipids staining them red

78
Q

First sudan dye to be introduced in histochemistry

A

Sudan III

79
Q

Sudan III is good for

A

fat stain for CNS tissues

80
Q

Sudan Black B, Sudan IV and Sudan III are considered what type of stain

A

Histochemical stain

81
Q

Other examples of histological stains

A 
Wrights
Giemsa
Methylene blue
Crystal violet
Carbol fuschin
82
Q

Accentuator is a part of staining reaction

True or False

A

False

83
Q

Components of acid alcohol

A

HCL + ethanol

84
Q

What is orthochromatic staining

A

Tissue stained with same shade/hue as the dye

85
Q

– uses aqueous or alcoholic dye solutions to produce a color

A

Direct Staining

86
Q

Metachromatic staining is used to stain what structures

A

Cartilage
Amyloid
Mast cell granules
Epithelial mucins

87
Q

2 examples of staining jar

A

couplin jar

slotted staining dishes

88
Q

Stains under Alum hematoxylin

A

Gills
Ehrlichs
Meyers
Coles

89
Q

Process of applying a mounting medium between the stained tissue section and the coverslip

A

mounting

90
Q

Reasons for doing mounting

A

permanent safekeeping
easy handling and storage
prevent bleaching of sections

91
Q

Mounting medium should have a refractive index close to that of the slide which is

A

1.518

92
Q

What is resinous mountant

A

dehydrated and cleared by xylene

permanent

93
Q

What is aqueous mountant

A

for water miscible preparation

semi-permanent, temporary

94
Q

Canada balsam’s refractive index is

A

1.524

95
Q

Routine mountant

A

Canada balsam

96
Q

Canada balsam is extracted from

A

Abus Balsamea

97
Q

Disadvantage of using canada balsam

A

Darkens with age

causes gradual fading of stains

98
Q

Enumerate types of resinous mountant and their refractive index

A

Canada balsam- 1.524
DPX- 1.532
XAM- 1.52
CLARITE- .544

99
Q

Glycerin jelly’s aka ______ and refractive index

A

Kaiser’s

1.47

100
Q

Farrant’s aka ______ and refractive index

A

Gum arabic

1.43

101
Q

Apathy’s refractive index

A

1.52

for methylene blue stained nerve preparations

102
Q

Brun’s is used for

A

frozen sections directly from water

103
Q

Water as mountant

A

not used because of its low refractive index

easily evaporates and cannot examine those under IOI

104
Q

Ringing is mandatory

True or False

A

False

105
Q

Sealing margins of coverslip

A

ringing

106
Q

Ringing is done to

A

prevent escape of fluid
prevent evaporation
prevent sticking of slides upon preparation
immobilize coverslip

107
Q

Type of ringing medium

A

Kronig cement

durofix

108
Q

Color that eosin stains

A

pink

109
Q

color that hematoxylin stains

A

blue

110
Q

examples of NEUTRAL / AMPHOTERIC DYE

A

Wrights
Irishman
Giemsa
Leishmann

111
Q

Process of coating slide with thin celloidin solutions. Done if sections will be subjected to strong alkali or acid solutions and for tissues containing glycogen for demonstration

A

COLLODIONIZATION

112
Q

Solvents Used for Stains

A

Water
Alcohol (methyl or ethyl)
Aniline water
Phenol

113
Q

Steps in RE-STAINING of OLD SECTIONS

A
  1. immerse in xylene for 24 hrs
  2. remover coverslip
  3. immerse in xylene for 30 mins
  4. immerse in 0.5% potassium permanganate for 5-10 mins. rinse with tapwater
  5. immerse in 5% oxalic acid
  6. re-stain
114
Q

Years of retention for Clinical pathology laboratory reports

A

10 yrs

115
Q

Years of retention for Autopsy forensic reports

A

indefinitely

116
Q

Years of retention for Surgical pathology and bone marrow Reports

A

10 yrs

117
Q

Years of retention for Cytogenetics report

A

20 yrs

118
Q

Years of retention for Pathology /Bone marrow slides

A

10 yrs

119
Q

Years of retention for Pathology blocks

A

10 yrs

120
Q

Years of retention for Cytogenetic slides

A

3 yrs

121
Q

Years of retention for Cytogenetic diagnostic images

A

20 yrs

122
Q

it governs the contrast between the cellular detail and the background, and also the transparency of the observed sample against the bright field of the microscope.

A

refractive index

123
Q

The mounting media must always have an RI _____ than the mounted sample to impart more transparency.

A

higher

124
Q

The slide may then be incubated at __________ after mounting, to harden the medium.

A

37 degC

12-24 hrs

125
Q

You can use immersion oil on an uncovered slide

True of False

A

False

126
Q

Oil can be applied, but do not attempt to wipe it off until the mounting medium is cured for how long?

A

at least overnight or in a hot plate or oven for 1 hr

127
Q

Do not store mounted slides vertically for ______ if cured at room temperature.

A

2 days

128
Q

Excessive mounting medium will cause it to ooze out of the sides of the cover glass, and should be carefully wiped with a fine cloth moistened with ____________.

A

xylene

129
Q

Excessive blotting will

A

dry up the section, causing shrinkage and cracking of the specimen.

130
Q

If the section has to be remounted, the cover glass may be removed by soaking in

A

xylene

131
Q

Excess xylene, if not removed, will

A

mix with the mountant and form bubbles on the slide.

132
Q

Too little mounting medium may

What is the remedy

A

cause improper setting of the coverslip or formation of bubbles on the section

teased out by gently pressing on the cover glass with a pointed forceps or mounting needle.

133
Q

Setting may be hastened in

A

hot oven at 50°C for 2 hours.

134
Q

basic aniline dyes should be mounted in

A

non-acid containing mountants

135
Q

Prussian blue reaction should be mounted in

A

non-reducing media

136
Q

a paper label should contain

A

patient’s name
section number
staining method used

137
Q

used for mounting sections from distilled water when the stains would be decolorized or removed by alcohol and xylene as would be the case with most of the fat stains (Sudan methods) or for metachromatic staining of amyloid.

A

AQUEOUS MOUNTING MEDIA

138
Q

glycerin jelly or gum arabic used to

A

solidify the medium

139
Q

glycerol used to

A

cracking and drying of the preparation

140
Q

sugar is used to

A

increase the refractive-index

141
Q

usually regarded as the standard mountant for fat stains.

A

glycerin

142
Q

standard mounting medium used when dehydration and clearing with xylene cannot be made

A

glycerin jelly

143
Q

recommended as an alternative for glycerine jelly.

A

Polyvinyl alcohol

144
Q

has the highest index of refraction

A

Pure glycerin

145
Q

used as a mountant in immunofluorescence microscopy

A

Polyvinyl alcohol

146
Q

The mountant is not set in the desired amount of hardness and therefore requires “ringing”

A

glycerin jelly

147
Q

does not solidify upon storage and therefore does not need to be heated before use.

A

gum arabic medium

148
Q

general purpose aqueous mountant

A

apathy’s

149
Q

not compatible with normal histological stains.

A

von apathy’s

150
Q

It is one of the most useful aqueous mountants for fluorescent microscopy, being virtually nonfluorescent.

A

apathy’s

151
Q

Canada balsam can be made neutral or acid by adding excess amounts of

A

calcium carbonate

salicylic acid

152
Q

It is a transparent, almost colorless oleoresin that adheres firmly to glass and sets to a hard consistency without granulation

A

Canada balsam

153
Q

substitute for xylene in canada balsam that may be less ofa health hazard but may cause other problems such as slow hardening and premature darkening.

A

histomount

154
Q

may be substituted for xylene as a solvent in canada balsam. The medium can only be neutralized temporarily since the mixture becomes acidic and changes into a brown color upon storage.

A

Benzene

155
Q

may be added to canada balsam to maintain its neutral reaction

A

calcium carbonate chips

156
Q

Canada balsam is recommended for

A

whole mounts and thick sections

157
Q

This is a resinous medium recommended for small tissue sections but not for whole mounts because of shrinkage produced on drying

A

DPX

158
Q

It has a greater advantage over Canada balsam in that slides can be cleaned of excess mountant simply by stripping it off after cutting around the edge of coverslip

A

DPX

159
Q

generally preferred over D.P.X

A

Clarite

160
Q

generally suitable for all enzymatic label/chromogen combinations and fluorescent labels.

A

Aqueous mounting medium

161
Q

quenches the staining intensity.

A

glycerol

162
Q

exposure to excitation light of most fluorescent labels which results in diminished staining

A

photo bleaching

163
Q

may form when the mounting media is too thin, and as it dries air is sucked in under the coverslip.

A

bubbles

164
Q

Contamination of clearing agents or cover slipping media may

A

produce a bubbled appearance under the microscope.

Histopathology Lec&Lab (7 decks)

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