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MTAP - HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES MODULE > LESSON 8: STAINING > Flashcards

LESSON 8: STAINING Flashcards

1
Q

 process of applying dyes on the sections to see and study the architectural pattern of the tissue and physical characteristics of the cells

A

Staining

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2
Q

 tissues and cells display varying affinities for most dyes and stains used during the process

A

Staining

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3
Q

 Acidic structures—greater affinity for
 Basic structures—greater affinity for

A

basic dyes

acidic dyes

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4
Q

 related procedure that makes use of heavy metal salts which are selectively precipitated on certain cellular and tissue components

A

Impregnation

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5
Q

 used for silver staining of nervous tissue and demonstration of reticulin

A

Impregnation

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6
Q

Impregnation  most commonly used agent:—may also be used as a staining agent

A

silver nitrate

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7
Q

THREE MAJOR TYPES OF STAINING

A
  1. HISTOLOGICAL STAINING
  2. HISTOCHEMICAL STAINING/HISTOCHEMISTRY
  3. IMMUNOHISTOCHEMICAL STAINING
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8
Q

METHODS OF STAINING

A
  1. DIRECT STAINING
  2. INDIRECT STAINING
  3. PROGRESSIVE STAINING
  4. REGRESSIVE STAINING
  5. METACHROMATIC STAINING
  6. COUNTERSTAINING
  7. METALLIC IMPREGNATION
  8. VITAL STAINING
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9
Q
  1. VITAL STAINING:
A

INTRAVITAL STAINING

SUPRAVITAL STAINING

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10
Q

 tissue constituents are demonstrated in sections by direct interaction with a dye or staining solution

A

HISTOLOGICAL STAINING

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11
Q

 producing coloration of the active tissue component

A

HISTOLOGICAL STAINING

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12
Q

 employed to demonstrate the general relationship of tissues and cells with differentiation of nucleus and cytoplasm

A

HISTOLOGICAL STAINING

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13
Q

 Examples: microanatomic stains, bacterial stains and specific tissue stains (muscles, CT, and neurologic stains)

A

HISTOLOGICAL STAINING

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14
Q

 tissue constituents are studied through chemical reactions that will permit microscopic localization of a specific tissue substance

A

HISTOCHEMICAL STAINING/HISTOCHEMISTRY

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15
Q

 Examples: Perl’s Prussian blue reaction for hemoglobin, Periodic Acid Schiff staining for carbohydrates

A

HISTOCHEMICAL STAINING/HISTOCHEMISTRY

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16
Q

 Enzyme histochemistry: active reagent serves as the substrate upon which the enzymes act

A

HISTOCHEMICAL STAINING/HISTOCHEMISTRY

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17
Q

 final opacity of coloration produced from the substrate rather than the tissue

A

HISTOCHEMICAL STAINING/HISTOCHEMISTRY

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18
Q

 combination of immunologic and histochemical techniques that allow phenotypic markers to be detected and demonstrated

A

IMMUNOHISTOCHEMICAL STAINING

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19
Q

 makes use of different labels: monoclonal/polyclonal, fluorescent-labeled, enzyme-labeled antibodies

A

IMMUNOHISTOCHEMICAL STAINING

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20
Q

 process of giving color to the sections by using aqueous or alcoholic dyes

A
  1. DIRECT STAINING
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21
Q

 process whereby action of dye is intensified by adding another reagent (MORDANT) which serves as a link/bridge between tissue and dye making staining reaction possible

A
  1. INDIRECT STAINING
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22
Q

may be applied to tissue before staining or may be included in the staining process, or may be incorporated as part of the dye solution itself

A

MORDANT

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23
Q

MORDANT Eg.

A

potassium alum with hematoxylin in Ehrlich’s hematoxylin and iron in Weigert’s hematoxylin

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24
Q

 not essential to the chemical union of tissue and dye

A

ACCENTUATOR

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25
Q

but merely accelerates or hastens the speed of staining reaction by increasing the staining power and selectivity of the dye

A

ACCENTUATOR

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26
Q

ACCENTUATOR Eg:

A

potassium hydroxide in Loeffler’s methylene blue and phenol in carbol thionine and carbol fuchsin

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27
Q

 process whereby tissue elements are stained in a definite sequence, and the staining solution is applied for specific periods of time or until the desired intensity of coloring of the different tissue elements is attained

A

PROGRESSIVE STAINING

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28
Q

 no decolorization or washing after the application of the dye

A

PROGRESSIVE STAINING

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29
Q

 tissue is first over stained to obliterate cellular details, and excess stain is removed or decolorized from unwanted parts of the tissue, until the desired intensity of color is obtained

A
  1. REGRESSIVE STAINING
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30
Q

: selective removal of excess stain from the tissue so that a specific substance may be stained distinctly from surrounding tissues

A

DIFFERENTIATION/DECOLORIZATION

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31
Q

 use of dyes which differentiate particular substances by staining them with a color that is different from that of the stain itself

A

METACHROMATIC STAINING

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32
Q

 basic dyes belonging to the thiazine and triphenylmethane groups

A

METACHROMATIC STAINING

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33
Q

METACHROMATIC STAINING Examples:

A

methyl violet or crystal violet Cresyl blue for reticulocytes Safranin Bismarck brown

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34
Q

stain tissues in color shades that are similar to the color of the dye itself

A

ORTHOCHROMATIC STAINING

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35
Q

 application of a different color or stain to provide contrast and background to the staining of structural components to be demonstrated

A

COUNTERSTAINING

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36
Q

Cytoplasmic stains:

A

Red eosin Y, Eosin B, Phloxine B
Yellow piciric acid, orange G, Rose Bengal
Green light green SF, Lissamine green

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37
Q

Nuclear stains:

A

Red neutral red, safranin 0, carmine, hematoxylin
Blue methylene blue, toluidine blue, Celestine blue

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38
Q

 process where specific tissue elements are demonstrated, not by stains, but by colorless solutions of metallic salts which are thereby reduced by the tissue, producing an opaque, usually black deposit on the surface of the tissue

A

METALLIC IMPREGNATION

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39
Q

 agent is not absorbed by the tissue, but is held physically on the surface as a precipitate or as a reduction product

A

METALLIC IMPREGNATION

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40
Q

 Examples: ammoniacal silver and gold chloride

A

METALLIC IMPREGNATION

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41
Q

reduced by argentaffin cells (melanin and intestinal glands) Gold chloride

A

Ammoniacal silver

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42
Q

 selective staining of living cell constituents, demonstrating cytoplasmic structures by phagocytosis of the dye particle

A

VITAL STAINING

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43
Q

**nucleus is resistant to staining

A

VITAL STAINING

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44
Q

***if nuclear structures are demonstrated > death of cell already

A

VITAL STAINING

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45
Q

 Examples: trypan blue>reticuloendothelial cells; Janus green>mitochondria

A

VITAL STAINING

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46
Q

 done by injecting dye into any part of the body (intravenous, intraperitoneal or subcutaneous), producing specific color

A

INTRAVITAL STAINING

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47
Q

 Examples: lithium, carmine and India ink

A

INTRAVITAL STAINING

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48
Q

 stain living cells immediately after removal from the living body

A

SUPRAVITAL STAINING

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49
Q

 Examples: neutral red, Janus green, trypan blue, etc.

A

SUPRAVITAL STAINING

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50
Q

 Regressive staining due to the decolorization step using acid alcohol

A

H and E Staining Technique

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51
Q

H and E Staining Technique  Major steps:

A

Deparaffinization (Step 1)
Hydration (Step 2)
Nuclear staining (Step 4)
Differentiation (Step 6)
Bluing (Step 8)
Counterstaining (Step 10)
Dehydration (Step 11)
Clearing (Step 12)

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52
Q

Nuclei:

A

blue to blue black

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53
Q

Karyosome:

A

dark blue

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54
Q

Cytoplasm, proteins in edema fluid:

A

pale pink

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55
Q

RBCs, eosinophilic granules, keratin:

A

bright orange-red

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56
Q

Basophilic cytoplasm, plasma cells, osteoblast:

A

purplish pink

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57
Q

Cartilage:

A

pink or light blue to dark blue

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58
Q

Calcium and calcified bone:

A

purplish blue

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59
Q

Decalcified bone matrix, collagen, osteoid:

A

pink

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60
Q

Muscle fibers:

A

deep pink

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61
Q

 extracted from the core/heartwood of a Mexican tree (Hematoxylin campechianum)

A

HEMATOXYLIN

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62
Q

 Ripening/Oxidizing: exposure to air & sunlight or to strong oxidizing agents

A

HEMATOXYLIN

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63
Q

Aluminum Hematoxylin:

Iron Hematoxylin:

Phosphotungstic Acid Hematoxylin:

A
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64
Q

Sodium iodate

A

Ehrlich’s

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65
Q

Tissues subjected to acid decalcification and acidic tissues

A

Ehrlich’s

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66
Q

Mercuric chloride

A

Harris’

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67
Q

Routine nuclear staining, exfoliative cytology and staining sex chromosomes

A

Harris’

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68
Q

Alcoholic iodine

A

Cole’s

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69
Q

Routine purposes, esp in sequence to Celestine blue

A

Cole’s

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70
Q

Sodium iodate

A

Mayer’s

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71
Q

Cytoplasmic glycogen

A

Mayer’s

72
Q

Ferric ammonium chloride

A

Weigert’s

73
Q

Muscle fibers and connective tissues

A

Weigert’s

74
Q

Ferric ammonium sulfate

A

Heidenhain’s

75
Q

Nuclear and cytoplasmic inclusions (chromatin, chromosomes, nucleoli, centrosomes, mitochondria)

A

Heidenhain’s

76
Q

Voluntary muscle striations and myelin

A

Heidenhain’s

77
Q

1% aq. phosphotungstic acid

A

Phosphotungstic Acid Hematoxylin

78
Q

Nuclei, fibrin, muscle striations, myofibrils, fibroglia

A

Phosphotungstic Acid Hematoxylin

79
Q

Collagen, bone, cartilage

A

Phosphotungstic Acid Hematoxylin

80
Q

Paraffin, celloidin and frozen sections

A

Phosphotungstic Acid Hematoxylin

81
Q

For demonstration of spermatogenesis

A

Copper Hematoxylin

82
Q

 Extracted from a female cochineal bug (Coccus cacti)

A

COCHINEAL DYES

83
Q

 Treated with alum to produce carmine

A

COCHINEAL DYES

84
Q

: neuropathological studies

A

Carmine + Picric acid > Picrocarmine

85
Q

: glycogen

A

Carmine + aluminium chloride > Best carmine

86
Q

 Vegetable dye from lichens treated with ammonia and exposed to air

A

ORCEIN

87
Q

 For staining elastic fibers: dark brown

A

ORCEIN

88
Q

 Coal tar dyes or aniline dyes

A

SYNTHETIC DYES

89
Q

: produces color

A

 Chromophores

90
Q

: promotes color retention

A

 Auxochromes

91
Q

 DYE:

A

Chromophore + auxochrome

92
Q

ACID DYES:
BASIC DYES:
NEUTRAL DYES:

A

Picric acid

Methylene blue

Romanowsky
Giemsa
Irishman

93
Q

Most sensitive

A

Sudan Black B

94
Q

Stains phospholipids and neutr1al lipids (BLACK)

A

Sudan Black B

95
Q

Recommended for staining neutral lipids (RED)

A

Sudan IV (Scharlach R)

96
Q

First to be introduced

A

Sudan III

97
Q

Fat stain for CNS tissues (ORANGE)

A

Sudan III

98
Q

Other Non-Lysochrome Lipid Stains:

A
99
Q

: for neutral fats and lipofuchsin (brilliant red)

A

 Oil Red O

100
Q

: stains lipids black

A

 Osmic Acid

101
Q

Most valuable for differential staining of CT & cytoplasm

A

Eosin

102
Q

Connective tissues

A

Acid Fuchsin-Picric Acid

103
Q

Discrimination between dead and living cells

A

Acridine Orange

104
Q

DNA: green fluorescence ; RNA: red fluorescence

A

Acridine Orange

105
Q

Calcium salts and sites of phosphatase activities

A

Acridine Red 3B

106
Q

Acid mucopolysaccharides

A

Alcian Blue

107
Q

Conterstaining of epithelial sections

A

Aniline Blue

108
Q

Deep staining of acid-fast organisms, mitochondria

A

Basic fuschin

109
Q

differentiation of smooth muscles

A

Basic fuschin With picric acid

110
Q

: detection of aldehydes

A

Basic fuschin In Feulgen’s and Schiff’s reagent

111
Q

: connective tissues, mucin and elastic tissues

A

Basic fuschin In Van Gieson’s

112
Q

Hemoglobin

A

Benzidine

113
Q

Contrast stain for Gram’s, acid fast and Pap’s method

A

Bismarck Brown

114
Q

Used for staining diphtheria

A

Bismarck Brown

115
Q

Chromatin stain for fresh materials (smear prep)

A

Carmine

116
Q

With aluminum chloride > glycogen

A

Carmine

117
Q

With aluminum chloride > glycogen

A

Carmine

118
Q

Routine staining of fixed sections

A

Celestine blue

119
Q

Axis cylinders in embryos

A

Congo Red

120
Q

: elastic tissues, amyloid and myelin

A

Congo Red In Krajians method

121
Q

Amyloid and platelets

A

Crystal violet

122
Q

Leukocyte differentiation

A

Giemsa

123
Q

Metallic impregnation

A

Gold sublimate

124
Q

Oldest stain

A

Iodine

125
Q

: microorganisms and fibrin

A

GRAM’S Iodine

126
Q

: glycogen, amyloid and corpora amylacea

A

LUGOL’s Iodine

127
Q

Mitochondria

A

Janus Green B

128
Q

Ascaris eggs, erythrocytes and bacterial spore

A

Malachite Green

129
Q

Chromatin

A

Methyl green

130
Q

Plasma cells

A

Methylene blue

131
Q

Cytologic examinations of fresh sputum for malignant cells

A

Methylene blue

132
Q

Nuclei of leukocytes

A

Methylene violet

133
Q

Observing cell granules and vacuoles in phagocytic cells

A

Neutral red

134
Q

Carbol fuchsin substitute in acid-fast staining

A

Night blue

135
Q

Elastic fibers (Taenzer Unna ? Method)

A

Orcein

136
Q

Dermatological studies to demonstrate finest and most delicate skin fibers

A

Orcein

137
Q

Stains fat

A

Osmium tetroxide

138
Q

Demonstration of circulatory system by injection

A

Prussian blue

139
Q

Fix and stain blood and glandular tissues

A

Rhodamine B

140
Q

Spirochetes, reticulum and other fiber stains

A

Silver nitrate

141
Q

Substitute for thionine in fresh frozen sections

A

Toluidine blue

142
Q

Recommended for Nissl granules or chromophilic bodies

A

Toluidine blue

143
Q

Neuroglia in frozen sections

A

Victoria blue

144
Q

Magenta Red

A
  1. Periodic Acid Schiff (PAS)
145
Q

Stain of choice for glycogen (red)

A

**PAS with Diastase Control

146
Q

Selective and specific for glycogen (bright red)

A
  1. Best Carmine
147
Q

Mahogany Brown

A
  1. Langhan’s Iodine
148
Q

Histones and protamines (green)

A
  1. Alkaline-fast Green
149
Q

Cystine and cysteine (blue-green)

A
  1. Peracetic acid-Alcian blue
150
Q

Arginine (orange-red)

A
  1. Sakaguchi’s Test (Milton’s Reagent)
151
Q

Most reliable and specific stain for DNA

A
  1. Feulgen Technique
152
Q

DNA: Red-purple

A
  1. Feulgen Technique
153
Q

DNA: Green or Blue-green
RNA: Rose-red

A
  1. Methyl Green-Pyronin
154
Q

Most widely-used fluorochrome

A
  1. Fluorescein
155
Q

Apple green

A
  1. Fluorescein
156
Q

Orange-red

A
  1. Rhodamine
157
Q

DNA: yellow-green
RNA: brick to orange-red

A
  1. Acridine Orange
158
Q

DNA: yellow

A
  1. Acriflavine
159
Q
  1. Gomori’s Silver Impregnation
A

Black

160
Q
  1. Van Gieson’s
A

Pink or Deep red

161
Q
  1. Masson’s Trichrome
A

Blue

162
Q
  1. Mallory’s Aniline
A

Blue

163
Q
  1. Weigert’s Elastic Tissue
A

Dark-blue or Blue-black

164
Q
  1. Taenzer-Unna Orcein
A

Dark brown

165
Q
  1. Verhoeff’s
A

Black

166
Q
  1. Krajian’s
A

Bright red

167
Q
  1. Perl’s Prussian Blue
A

Deep blue

168
Q
  1. Masson Fontana
A

Black

169
Q
  1. Von Kossa’s Silver Nitrate
A

Black

170
Q
  1. Levaditi’s
A

Black

171
Q
  1. Warthin-Starry Method
A

Black

172
Q
  1. Wade-Fite Technique
A

Red

173
Q
  1. Orcein Method
A

Brown-black

174
Q

also used to remove stains on the skin

A

1% acid alcohol

175
Q

Other Bluing Agents:

A

warm tap H2O: 40-50degC, bicarbonate, potassium/sodium acetate, ammonia and Scott’s Tap Water Substitute (TWS)

176
Q

more specific for connective tissue and epithelial mucin

A

Alcian blue

177
Q

more specific for connective tissue and epithelial mucin

A

Alcian blue

MTAP - HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES MODULE (9 decks)

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