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HISTOPATH > LECT | Staining FINALS > Flashcards

LECT | Staining FINALS Flashcards

1
Q

Enables one to see and study the architectural pattern of the tissue and physical characteristics and structural relationships of tissues and their cells

A

Staining/Dyeing

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2
Q

Staining/Dyeing is made possible through:

A
  1. Capillary osmosis
  2. Solubility
  3. Absorption
  4. Adsorption of stains or dyes by tissues
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3
Q

Alkaline structures (cytoplasm) are stained by

A

acidic stains

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4
Q

Eosin stains cytoplasm color?

A

Red

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5
Q

Acidic structures (nucleus) are stained

A

alkaline/basic stain

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6
Q

hematoxylin stains nucleus color?

A

Blue

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7
Q

Classification of Staining

A
  1. Histological staining
  2. Histochemical Staining (Histochemistry)
  3. Immunohistochemical Staining
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8
Q
  1. The process whereby the tissue constituents are demonstrated in section by direct interaction with a dye or staining solution, producing coloration of the active tissue component.
  2. Used to demonstrate the general relationship of tissues and cells with differentiation of nucleus and cytoplasm.
A

Histological Staining

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9
Q

Histological Staining examples

A
  1. Microanatomic stains
  2. Bacterial stains
  3. Specific tissue stains (muscles, connective tissue, neurologic stains)
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10
Q

The process whereby various constituents of tissues are studied through chemical reactions that will permit microscope localization of a specific tissue substance

A

Histochemical Staining or Histochemistry

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11
Q

Histochemical Staining or Histochemistry example

A
  1. Perl’s Prussian Blue (hemoglobin); demonstrates ferric ion (Fe3+) in tissues
  2. Periodic Acid Schiff (carbohydrates)
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12
Q

A combination of immunologic and histochemical techniques that allow phenotypic markers to be detected and demonstrated under the microscope, using a wide range of monoclonal, fluorescent-labeled or enzyme- labeled antibodies.

A

Immunohistochemical Staining or Immunohistochemistry

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13
Q
  1. It is the basis of histochemistry
  2. It is accomplished by controlled, specific chemical reactions designed to give a final color (staining) at the site/location of the structure of the substance in the cells or tissue
A

Specific Staining

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14
Q

Specific Staining example

A
  1. Perl’s Prussian blue reaction – Hemosiderin
  2. Periodic Acid Schiff technique – Polysaccharides
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15
Q

The staining of the tissue by means of simple alcoholic/aqueous solution of the dye.

A

Simple Staining/Direct Staining

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16
Q

Simple Staining/Direct Staining examples

A

Methylene blue and Eosin

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17
Q

The action of the dye is intensified by some other agents.

  1. Mordant
  2. Accentuator
A

Indirect Staining

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18
Q
  1. Substance which when taken up by the tissue, helps make the in return serving as a link or bridge to make the staining reaction possible
  2. Combines with a dye forming a colored “lake” which combined with tissues to form an insoluble “tissue- mordant-dye complex”
  3. An integral part of the staining reaction itself, without which, no staining could possibly occur
A

Mordant

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19
Q

Indirect staining Mordant example

A
  1. Potassium Alum with Hematoxylin in Erlich’s hematoxylin
  2. Iron in Welgert’s hematoxylin
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20
Q
  1. Chemical substances that does not participate but merely increases or heightens the color intensity, selectively and crispness of the stain
  2. Differ from mordants in that they do not bind or link the tissue to the dye
A

Accentuator

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21
Q

Indirect staining Accentuator example

A
  1. Potassium hydroxide (KOH) in Loeffler’s Methylene Blue
  2. Phenol in carbol thionine and carbol fuchsin
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22
Q
  1. Staining is continued in a definite sequence until the desired intensity of coloring of the different tissue elements is attained.
  2. No washing/differentiation/decolorization in between is required, solely relies on the selective affinity of dyes for different cellular elements
A

Progressive Staining

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23
Q

Any stain is possible as long as no differentiation is done

A

Progressive Staining

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24
Q

The tissues are overstained and the excess dye is then removed until the desired intensity is obtained

A

Regressive Staining

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25
Q

Regressive Staining : 2 principle

A
  1. Overstaining
  2. Washing/ Differentiation/ Decolorization
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26
Q

Example of Regressive staining

A
  1. Acid Fast Staining
  2. Gram Staining
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27
Q

Entails the use of the specific dyes that stains tissues with a color that is different from that of the stain color itself

A

Metachromatic Staining

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28
Q

Metachromatic Staining is employed for staining ?

A
  1. Cartilage
  2. Epithelial mucins
  3. Connective tissues
  4. Amyloid
  5. Mast cell granules
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29
Q

Metachromatic dye – basic dyes belonging to what group

A

Thizine and Triphenylmethane groups

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30
Q

Examples of Metachromatic Stain

A

Cresyl blue for reticulocytes

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31
Q

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

A

Counterstaining

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32
Q

Examples of Counterstain

A

Eosin ⎯ for cytoplasm

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33
Q
  1. The selective staining of living cells constituents, demonstrating cytoplasmic structures by phagocytosis if dye particle (cytoplasmic phagocytosis)
  2. The nucleus of the living cell is resistant to vital stains, and therefore is not demonstrated
  3. Demonstration of nuclear structures during vital staining suggests permeability of the membrane by the dye, signifying the death of the cell.
A

Vital Staining

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34
Q

Trypan blue and Janus Green B are examples of what staining classification

A

Vital Staining

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35
Q

Vital stain of reticuloendothelial cells

A

Trypan blue

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36
Q

true vital staining of mitochondria

A

Janus Green B

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37
Q

Done by injecting the dye into any part of the animal body (either intravenous, intraperitoneal, or subcutaneous) producing specific coloration of certain cells, particularly those of the RES.

A

Intravital Staining

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38
Q

Intravital Staining : commonly used dye

A
  1. Lithium
  2. India Ink
  3. Carmine
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39
Q

Used to stain living cells immediately after the removal from the living body

A

Supravital Staining

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40
Q

Example of supravital stain - probably the best vital dye

A

Neutral RED

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41
Q
  1. Makes use of heavy metals which are precipitated with selectivity of certain cellular and tissue components
  2. Has its greatest application in tissue from the CNS and for the demonstration of reticulin
  3. Differs from staining because it consists of an opaque black particulate precipitate
A

Metallic Impregnation

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42
Q

Metallic Impregnation examples

A
  1. Silver nitrate
  2. Osmium tetroxide
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43
Q

most commonly used agent for impregnation

A

Silver Nitrate

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44
Q

used for the demonstration of lipids

A

Osmium tetroxide

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45
Q

Are those obtained from plants and animal, previously utilized for dyeing of wool and cotton

A

Natural Dyes

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46
Q

most important and the most commonly dye used for routine histologic studies

A

Hematoxylin

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47
Q
  1. A vegetable dye extracted from lichens which are normally colorless, but when treated with ammonia and expose to air, produce blue or violet colors.
  2. Mainly used for staining Elastic fibers.
A

Orcein

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48
Q

An old histologic dye extracted from the female cochineal bug (coccus cacti)

A

Cochineal

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49
Q

A plant with orange stigmas yielding a dye

A

Saffron

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50
Q

Examples of natural dyes

A
  1. Hematoxylin
  2. Orcein
  3. Cochineal
  4. Saffron
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51
Q

Sometimes known as “coal tar dyes” since they were originally manufactured from substances that have been taken from coal tar

A

Synthetic Dyes

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52
Q

Synthetic Dyes consist of substances

A
  1. Chromophores
  2. Auxochromes
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53
Q
  1. Greek.“color-bearers”
  2. Substances capable of producing visible colors
A

Chromophores

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54
Q
  1. Greek.“color-increasers”
  2. Substances which impart to the compound property of electrolytic dissociation, thereby altering the shade of the dye, giving it the property of forming salts with another compound and ultimately retaining its color.
  3. The introduction of a chromogenic group into a uncolored molecule will cause it to be colored, it will then be a chromogen, which is colored and not a dye
A

Auxochromes

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55
Q

Synthetic dye coloring property is attributed to the

A

chromophore

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56
Q

Synthetic dye, which dyeing property to the salt forming

A

Auxochrome

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57
Q
  1. A process of selective removal of excess dye
  2. If the dye is a basic one, differentiation is carried out by an acid solution
  3. An alkaline medium is used for differentiation after an
    acid dye
A

Differentiation

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58
Q
  1. Natural dye is extracted from the core of heartwood of the tree Hematoxylin campechianium
  2. By far the most valuable staining reagent used by the cytologist due to its powerful nuclear and active coloring agent
A

Hematoxylin

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59
Q

Hematin

A

formed by oxidation of Hematoxylin RIPENING

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60
Q
  1. Usually done by exposing the substances to air and sunlight, thereby oxidizing this
  2. A slow process (3-4months) but it can be accelerated by adding strong oxidizing agents
A

Hematoxylin

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61
Q

complex of stain and mordant is called a

A

“lake”

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62
Q

Mordant example of Hematoxylin

  1. most commonly used mordant. Gives a blue take appearance
  2. Increase the selectivity for nuclei, especially if acid is added or is used as a differentiating agent
A

Aluminum

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63
Q

Mordant example of hematoxylin:

caused an intense blue-black appearance when used as a mordant

A

Ferric salt

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64
Q

Hematoxylin: mordant example

A
  1. Aluminum
  2. Chromium
  3. Iron
  4. Copper
  5. Tungsten/Phosphotungtic acid (PTAH)
  6. Ferric salt
65
Q
  1. A red acid dye used for cytoplasm differentially
  2. Commonly used as a background or contrast stain because it gives a leasing and useful contrast to nuclear stains such as hematoxylin
A

Eosin

66
Q

examples of Eosin

A
  1. Eosin Y
  2. Eosin B
  3. Eosin S/Ethyl Eosin
67
Q

Most common eosin

A

Eosin Y

68
Q

H and E Staining Color Reaction:

  1. Cytoplasm & proteins in edema fluid
A

Pale pink

69
Q

H and E Staining Color Reaction:

  1. Decalcified bone
  2. Osteoid
  3. Collagen
A

Pink

70
Q

H and E Staining Color Reaction:

Muscle fibers

A

Deep pink

71
Q

H and E Staining Color Reaction:

RBC
Eosinophil granules
Keratin

A

Bright orange to Red

72
Q

H and E Staining Color Reaction:

Calcium & Calcified bone

A

Purplish blue

73
Q

H and E Staining Color Reaction:

  1. Plasma cells
  2. Osteoblast
  3. Basophilic cytoplasm
A

Purplish pink

74
Q

H and E Staining Color Reaction:

Karyosome

A

Dark blue

75
Q

H and E Staining Color Reaction:

Nucleus

A

Blue to blue-black

76
Q

H and E Staining Color Reaction:

Cartilage

A

Light blue to Dark blue

77
Q
  1. A specialized fixative used in frozen section which serves to localize antigen and hydrolytic enzyme
  2. It is also used for preservation of lipids
A

Formol Calcium

78
Q
  1. It is similar with H and E technique but the differentiation stage is omitted
  2. It is a progressive form of staining
A

Modified H and E Technique

79
Q

Used for determining cervical cancer

A

Papanicolaou stain

80
Q

Papanicolaou stain makes use of 3 stains

A
  1. Hematoxylin
  2. OG-6 (orange green)
  3. Eosin Azure
81
Q

For staining cytoplasm of mature cells.
Example: mature superficial cells

A

OG-6 (orange green)

82
Q

for staining cytoplasm of immature cells.
Example: Intermediate and parabasal cells

A

Eosin Azure

83
Q

Eosin Azure composition?

A
  1. Eosin
  2. Bismarck brown
  3. Lithium carbonate
  4. PTA
  5. Light green stain (36,50,65)
84
Q
  1. It omitted the Bismarck brown dye for the EA solution
  2. Sharpness of color and brilliant staining reactions are improved
A

Modified Pap’s Stain

85
Q
  1. Mixture of picric acid and acid fuchsin
  2. for the demonstration of connective tissues, mucin and elastic tissue
A

Van Gieson’s (Acid fuchsin picric acid)

86
Q

Basic acridine fluorochrome which permits discrimination between dead and living cells

A

Acridine Orange

87
Q

Acridine orange gives this fluorescence color for

  1. DNA
  2. RNA
A
  1. GREEN fluorescence for DNA
  2. RED fluorescence for RNA
88
Q

For calcium salts and phosphate activity

A

Acridine Red 3B

89
Q
  1. Stain acid mucopolysaccharide
  2. More specific for connective tissue and epithelial mucin
A

Alcian blue

90
Q

A cytoplasmic stain used for counterstaining of epithelial sections

A

Aniline blue

91
Q

A plasma stain utilized also for deep staining of acid fast organisms mitochondria, and differentiation of smooth muscles with the use of picric acid

A

Basic Fuchsin

carbol fuchsin, coleman’s Feulgen, schiff’s, Mallory’s fuchsin, Aldehyde fuchsin (Gomori’s Stain)

92
Q

Used for staining hemoglobin

A

Benzidine

93
Q
  1. Used for staining diphtheria
  2. Used for contrast stain for gram’s technique, acid Papanicolaou method fast and Papanicolaou method
A

Bismarck brown

94
Q
  1. Used as a chromatin stain for fresh materials in smear preparations
  2. combined with aluminum chloride to stain glycogen
A

Carmine

95
Q

A mordanted dye acting as a basic dye and staining acid substances

A

Mayer’s Carmalum solution

96
Q

Recommended for routine staining of fixed sections

A

Celestine blue

97
Q
  1. Best known as indicator
  2. May be utilized as a stain for axis cylinders in embryos
  3. Used for staining tissues, amyloid and myelin (Krajian’s method)
A

Congo Red

98
Q

A nuclear or chromatin stain for staining amyloid in frozen sections and platelets

A

Crystal violet

99
Q

mixture of crystal violet, methyl violet in frozen sections

A

GENTIAN VIOLET

100
Q

Used for staining blood to differentiate leukocytes

A

Giemsa

101
Q

Used for metallic impregnation, made up of gold chloride and mercuric chloride

A

Gold Sublimate

102
Q
  1. The oldest of all stains
  2. Stains amyloid, cellulose, starch, carotenes, and glycogen.
A

Iodine (gram’s, lugol’s)

103
Q

Used for demonstrating mitochondria (intravital staining)

A

Janus Green B

104
Q
  1. Used as a contrast for staining ascaris and erythrocytes
  2. Used as bacterial spore staining
A

Malachite Green

105
Q

Stains chromatin green in the presence of an acid

A

Methyl Green

106
Q
  1. Plasma cells, fresh sputum for malignant cells, evaluation, and differentiation of bacterial organism.
  2. Diphtheria diagnosis and nervous tissue vital staining
A

Methylene Blue

107
Q

Used in frozen sections for rapid diagnosis

A

Loeffler’s Polychrome Methylene Blue

108
Q

Coloring nuclei of leukocytes reddish-purple in presence of methylene blue

A

Methylene violet

109
Q

For observing cell granules an vacuoles of phagocytic cells

A

Neutral red

110
Q

Substitute for carbol fuchsin in acid- fast staining

A

Night Blue

111
Q
  1. An excellent stain for elastic fibers
  2. Demonstrates the finest and most delicate fiber skin (dermatological studies)
A

Orcein

112
Q

Used as a fixative Used to stain fats

A

Osmium tetroxide

113
Q
  1. Normally utilized for the manufacture of paints
  2. Used as microanatomical contrast for demonstration for the circulatory system by injection (intravital staining)
A

Prussian blue

114
Q

Used with osmic acid to fix and stain blood and glandular tissues

A

Rhodamine B

115
Q

Used in identification of Spirochetes, reticulum and other fiber stains

A

Silver Nitrate

116
Q

Recommended for staining of Nissl granules or chromophilic bodies; nuclear stain for fixed tissues

A

Toluidine Blue

117
Q

Demonstration of neuroglia in frozen sections

A

Victoria Blue

118
Q

RED Cytoplasmic Counterstains

A
  1. Eosin Y
  2. Eosin B
  3. Phloxine B
  4. Rose Bengal
119
Q

YELLOW Cytoplasmic Counterstains

A
  1. Picric acid
  2. Orange G
120
Q

GREEN Cytoplasmic Counterstains

A
  1. Light green Sf
  2. Lissamine green
121
Q

RED Nuclear Counterstains

A
  1. Neutral red
  2. Safranin
  3. Carmine
122
Q

BLUE Nuclear Counterstains

A
  1. Methylene blue
  2. Toluidine blue
  3. Celestine blue
  4. Hematoxylin
123
Q

What are the Lysochromes (Oil Soluble Dyes)

A
  1. Sudan Black B (SBB)
  2. Sudan IV or Scharlach R
  3. Sudan III
124
Q
  1. Greatest affinity for phospholipids or neutral fats (triglycerides)
  2. A more sensitive coloring agent than other lysochromes
  3. Should be discarded if the brownish black color appeared
  4. Demonstrates lipids that are resistant to paraffin embedding
A

Sudan Black B (SBB)

125
Q
  1. Recommendedforneutralfats(triglycerides)
  2. Do not color phospholipids and fine lipid droplets
A

Sudan IV or Scharlach R

126
Q
  1. First Sudan Dye introduced into Histochemistry
  2. Fat soluble
  3. A good stain for the CNS
A

Sudan III

127
Q
  1. Used in order to promote adhesion of sections to slides
  2. Spread thinly and evenly on a clean grease-free glass slide which is then gently approximated to the end of the ribbon and drawn upwards in a near vertical motion
A

ADHESIVES

128
Q

Mayers’s Egg Albumin Formula:

A
  1. Egg white (50 mL)
  2. Glycerin (50 mL)
  3. Thymol crystals (100 mg)
129
Q

Dried Albumin Formula

A
  1. Dried albumin (5g)
  2. Sodium chloride (5g)
  3. Distilled water (100 mL)
  4. Thymol crystals (2 g)
130
Q

1% Gelatin Formula

A
  1. Gelatin (1g)
  2. Distilled water (100 mL)
  3. Glycerol (15 mL)
  4. Phenol crystals (2g)
131
Q

Gelatin Formula:

A
  1. 1% Gelatin (5 mL)
  2. 2% Formaldehyde (5 mL)
132
Q

Starch paste Formula:

A
  1. Powdered starch (1g)
  2. Distilled water (30 ml = 10 mL cold, 20 mL boiling)
133
Q

Readily available from outdated blood stored in blood banks, dispensed into sterile tubes of 0.5 ml each

A

Plasma

134
Q
  1. 0.1% aqueous detergent solution which is further diluted 1:10 with distilled water prior to use (final dilution: 0.01%)
  2. Used as section adhesive in immunochemistry
A

Poly-L-Lysine

135
Q

Very useful in cytology particularly for cytospin preparation if proteinaceous or bloody material

A

3-APES (3-aminopropyltriethoxysilane)

136
Q
  1. A syrupy fluid applied between the section and the coverslip after staining setting the section firmly, preventing the movement of the coverslip
  2. Necessary to protect the stained section from physical injury and from bleaching or deterioration of the stain as a result of oxidation
A

Mounting

137
Q

In mounting, for best results with stained tissue sections, they must be impregnated by an transparent medium having a refractive index close to that of

A

91.518

138
Q

2 groups of mounting media

A
  1. Aqueous / Water Soluble Mountants
  2. Resinous Mountants
139
Q

Designed to mount water-miscible preparations directly water in cases where the stain is removed or decolorized with alcohol or xylene

A

Aqueous / Water Soluble Mountants

140
Q
  1. Low refractive index
  2. Good only for temporary mounting
A

Water

141
Q

Mountant:
1. Also used as preservative
2. Has high index of refraction

A

Glycerin

142
Q

Glycerin RI

A

RI: 1.47

143
Q

Mountant:

  1. Does Not solidify upon storage
A

Farrant’s medium (Gum syrup)

144
Q

Farrant’s medium (Gum syrup) RI

A

RI: 1.43

145
Q

Used for methylene blue-stained nerve preparations and as general purpose aqueous mountant

A

Apathy’s medium

146
Q

Recommended for mounting frozen sections from water or paraffin sections which require dehydration and clearing

A

Brun’s fluid

147
Q

Examples of Aqueous / Water Soluble Mountants

A
  1. Water
  2. Glycine
  3. Farrant’s medium or gum syrup
  4. Apathy’s medium
  5. Brun’s fluid
148
Q

Used for preparations that have been dehydrated and cleared in xylene or toluene and are recommended for majority of staining methods.

A

Resinous Mountants

149
Q

Examples of Resinous Mountants

A
  1. Canada Balsam
  2. DPX
  3. XAM
  4. Clarite
150
Q
  1. natural resin extracted from the Canadian Tree (Abus balsamea)
  2. Recommended for whole mounts and for thick sections because it does not shrink
  3. Miscible with xylene and is quite expensive
A

Canada Balsam

151
Q

Canada Balsam RI

A

RI: 1.524

152
Q

Mountant recommended for small tissue sections

A

DPX

153
Q

DPX RI

A

RI: 1.532

154
Q

Synthetic resin mixture in xylene in pale yellow or colorless solution

A

XAM

155
Q

XAM RI

A

RI: 1.52

156
Q

Synthetic resin which is soluble in xylene (usually diluted to 60% with xylene)

A

Clarite

157
Q

Clarite RI

A

RI: 1.544

158
Q

2 mounting media most commonly used today

A
  1. Eukitt
  2. Entellan

HISTOPATH (4 decks)

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