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HPCT > Staining > Flashcards

Staining Flashcards

1
Q

The process of applying dyes on the sections to see and study the __________ of the tissue and _______ _______ of the cells

A

Staining

architectural pattern

physical characteristics

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2
Q

FACTORS INFLUENCING DYE BINDING

A

pH of the sol’n
INCREASE temp
INCREASE conc. of dye molecules
[+] other salts
Fixative type

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3
Q

Staining of Tissues can be Classified into 3 Major Groups

A

Histological staining
Histochemical staining (Histochemistry)
Immunohistochemical staining

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4
Q

The process whereby tissue constituents are demonstrated in sections by direct interaction with a dye

A

Histological staining/Microanatomical staining

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5
Q

used to demonstrate the general relationship of tissues and cells with differentiation of _________

A

Histological staining/Microanatomical staining

nucleus cytoplasm

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6
Q

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

A

Histochemical staining/Histochemistry

microscopic

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7
Q

Example of dyes whereby various constituents if tissues are studied through CHEMICAL RXN that will permit MICROSCOPIC LOCALIZATION of a SPECIFIC tissue substance

A

Per’s prussian blue rxn for hemoglobin

Periodic Acid Shiff staining for carbohydrates

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8
Q

__________
allow phenotypic markers to be detected and demonstrated under the microscope, using a wide range of ff Antibodies: [4]

A

Immunohistochemistry

Polyclonal
Monoclonal
Fluorescent labeled
Enzyme-labeled

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9
Q

The process by W/O sections are stained with simple Aqueous or Alcoholic solutions of the dye

A

Direct Staining

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10
Q

The process by which sections are stained with an alcoholic solutions of dye with the use of a mordant/accentuator

A

Indirect Staining

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11
Q

A substance that serves as a link or bridge to make the staining reaction possible.

What do we call the complex

A

Mordant

tissue-mordant-dye complex

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12
Q

Stains that serves as as a link or bridge to make the staining reaction possible.

A

Potassium Alum w/ Hematoxylin in Erlich’s hematoxylin

Iron in Weidgert’s hematoxylin

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13
Q

___________
A substance that does not participate in staining reaction, but merely accelerate or hasten the speed of staining reaction by increasing the [2]

A

Accentuator

Staining power
Dye Selectivity

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14
Q

Accentuators that does not participate in staining reaction, but merely accelerate or hasten the speed of staining reaction by increasing the staining power and selectivity of the dye

A

Potassium Hydroxide in Loeffler’s Methylene Blue
Phenol in Cabon Thionin
Carbol Fuschin

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15
Q

➔ Stained in definitive sequence
➔ Not washed / decolorized

A

Progressive staining

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16
Q

➔ Differentiation of tissue details relies solely on the
SELECTIVE AFFINITY of dyes for different cellular elements

A

Progressive staining

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17
Q

A progressive stain that relies solely on the
SELECTIVE AFFINITY of dyes for different cellular elements

A

Papanicolau smear

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18
Q

The tissue is first over stained to obliterate the cellular details, and then removed or decolorized from unwanted parts of the tissue

A

Regressive staining

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19
Q

Selective removal of excess stain from the tissue during regressive

A

Differentiation/Decolorization

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20
Q

➔ Entails the use of specific dyes which differentiate particular substance by staining them with a color that is different from that of the stain itself.

A

Metachromatic staining

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21
Q

Metachromatic dyes are basic dyes belonging to [2]

A

Thizine group

Triphenylmethane group

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22
Q

Metachromatic staining are designated for these locations [5]

A

Cartilage
CT
Epithelial mucins
Mast cell granules
Amyloid

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23
Q

Basic dyes belonging to the thizine and triphenylmethane groups [9]

A

Azure A,B,C
Basic fuschin
Bismarck brown
Cresyl Blue
Methyl Violet
Methylene Blue
Safranin
Thionine
Toluidine blue

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24
Q

Used to demonstrate the general relationship of tissues and cells with general differentiation of nucleus and cytoplasm without necessarily emphasizing the inclusion bodies

A

Microanatomical staining

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25
Q

Specific tissue elements are demonstrated by colorless solutions of metallic salts.

A

Metallic Impregnation

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26
Q

A stain example where specific tissue elements are demonstrated by colorless solutions of metallic salts.

A

Ammoniacal Silver – produces black

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27
Q

_________
➔ Different from a stain. It is absorbed by the tissue, but held [chemically/physically] on the surface as a ________ or as a _______ product in certain tissue components.
➔ [COLORS]

A

Metallic impregnation

physically

Precipitate
Reduction product

Gold/Silver

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28
Q

_____________
A selective staining of living cell constituents, demonstrating cytoplasmic structures by _______ of the dye particle

A

Vital staining

phagocytosis

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29
Q

Examples of vital stains [+designation]

A

Typan Blue – RE system
True Vitals Staining- mitochondria

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30
Q

Staining of living cells is done by injecting the dye into any parts of the animal body

A

Intravital staining

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31
Q

Stains/Dyes that stains living cells by injecting the dye into any parts of the animal body

A

Lithium
India ink
Carmine

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32
Q

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

A

Supravital staining

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33
Q

Common Vital Stain/Dyes

A

Neutral Red
Janus Green
Trypan Blue

Nile Blue
Thionine
Toluidine Blue

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34
Q

Best vital dye

A

Neutral red

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35
Q

Recommended for mitochondria

A

Janus Green

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36
Q

Toxin when stand for more than 1 hour

A

Trypan Blue

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37
Q

Enumerate all methods of staining [11]

A

Direct
Indirect
Progressive
Regressive
Differentiation/Decolorization
Metachromatic staining
Microanatomical staining
Metallic Impregnation
Vital
Intravital
Supravital

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38
Q

Categories of Staining [2]

A

Natural
Synthetic

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39
Q

Obtained from plants and animals previously dyeing of wool and cotton

A

Natural dyes

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40
Q

Examples of natural dyes that were obtained from plants and animals previously dyeing of wool and cotton

A

Cochineal dyes [+derivatices]
Orcein
Saffron
Hematoxylin

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41
Q

dye recommended for mitochondria

A

Janus green

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42
Q

becomes toxin when stand for more than 1 hr

A

Trypan blue

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43
Q

Natural dye derived by either extraction from the heartwood of a _______ tree
➔ Hematoxylin ________ (B.E.Q.)

A

Hematoxylin

mexican

campechianum

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44
Q

Considered as the valuable staining reagent
by cytologist

A

Hematoxylin

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45
Q

➔ Has powerful nuclear and chromatin staining
capacity

A

Hematoxylin

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46
Q

➔ Has striking polychrome properties

A

Hematoxylin

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47
Q

_________
Old histologic dye derived from extract female _____ [other term] -
(B.E.Q)

A

Cochineal dyes

cochineal bug
coccus cacti

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48
Q

Cochineal dyes are treated with _______ or produce a dye known as _______.

A

alum

carmine

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49
Q

extensively used as powerful chromatin and nuclear stain for fresh material and smear preparation

A

Cochineal dyes

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50
Q

_________
Vegetable dye extracted from certain ______ (B.E.Q.)

A

Orcein

lichens

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51
Q

Normally colorless but when treated with ______ and expose to _____ produce [2 colors]

A

ammonia

air

blue/violet

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52
Q

a dye that is weak acid & soluble in alkali

A

Orcein

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53
Q

Mainly used for elastic fibers

A

Orcein

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54
Q

_________
Derived from the fried stigmata of ________ (B.E.Q.)

A

Saffron

croccus sativus

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55
Q

Known as “coal tar dyes” as they are manufactured from substance that have been taken from coal tar

A

Synthetic dyes

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56
Q

Synthetic dyes are derived from

A

hydrocarbon benzene

57
Q

Synthetic dyes are collectively known as

A

Aniline dyes

58
Q

Synthetic dyes are classified into 3 groups

A

Acid dyes
Basic dyes
Natural dyes

59
Q

Coloring substance are found in the acid component

A
60
Q

an example of acid dyes that is outstanding in the sense that it is the only substance so far that can fix the stain tissues all by itself

A

Picric acid

61
Q

Acid radical usually taken from sulfuric or hydrochloric acid

A

Basic dyes

62
Q

an example of a basic nuclear stain; may be used both as an indicator and as a dye

A

Methylene blue

63
Q

Formed by combining aqueous solution of acid and basic dyes

A

Natural dyes

64
Q

Capable of staining cytoplasm and nucleus simultaneously and differentially

A

Natural dyes

65
Q

examples of natural dyes that are for leukocyte differentiation

A

Giemsa stain
Leishmann’s stain

66
Q

One of the principal tissue stains used in histology.

A

H&E stain

67
Q

most widely used stain in medical diagnosis and is often the gold standard

A

H&E stain

68
Q

H&E stain is the combination of 2 histological stains: [2]

A

Hematoxylin
Eosin

69
Q

The hematoxylin stains [cell part] [color] and eosin stains the [cell part] [color], with other structures taking on different shades, hues, and combinations of these
colors.

A

nuclei
-blue

EM
cytoplasm
-pink

70
Q

The stain shows the general layout and distribution of cells and provides a general overview of a tissue sample’s structure. Hence a pathologist can easily differentiate
between the nuclear and cytoplasmic parts of a cell.

A

H&E staining

71
Q

A natural dye derived from extraction from the heartwood of the mexican tree known as “hematoxylin campechianum”

The tree is grown commercially in ________.

The first person to use hematoxylin in histology
was ________ in [Date]

A

Hematoxylin

Jamaica

Wadeyerl
1862

72
Q

Progressive staining unlike in routine type which is a regressive staining because we use _________.

A

Modified H&E Staining

73
Q

A type of staining:
➔ No differentiation phase
➔ Used in frozen section

A

modified h&e staining

74
Q

May be done by exposing the substance to air and sunlight (slow)

A

Ripening/Oxidation

75
Q

Ripening/Oxidation may be done by adding oxidizing agents such as

A

Hydrogen peroxide
Mercuric oxide
Potassium permanganate
Sodium perborate
Sodium iodate

76
Q

dangerous if dry

A

picric acid

77
Q

Types of hematoxylin [4]

A

Alum hematoxylin
Iron Hematoxylin
Tungsten Hematoxylin
Copper Hematoxylin

78
Q

Used in routine H and E

A

Alum hematoxylin

79
Q

Mordant in alum hematoxylin

A

Potash alum/ Potassium aluminum sulfate

80
Q

_____________
produce good nuclear stain [color]

A

Alum Hematoxylin

red

81
Q

Dye examples that are slowly ripened

A

Erlich’s
Delafield’s

82
Q

Dye examples that are w/ oxidizing agent: sodium iodate

A

Mayer’s
Gill’s

83
Q

Dye examples that are w/ oxidizing agent: mercuric oxide

A

harris

84
Q

In Iron Hematoxylin,
________are used as oxidizing agents and mordant

A

Iron salts

85
Q

Examples under Iron Hematoxylin

A

Weigert’s

Heidenhain’s Susa

86
Q

a type of iron hematoxylin stain that is a ferric chloride

A

Weigert’s

87
Q

Weigert is in combination with _________ stain, can
demonstrate_______ elements and
___________ sections

A

Van gieson’s

CT
Entamoeba histolytica

88
Q

a type of iron hematoxylin stain that is for muscles/CT fibers

A

Weigert’s

89
Q

Standard iron hematoxylin

A

Weigert’s

90
Q

Van gieson’s stain is good for demonstrating ______.

A

collagen

91
Q

a type of iron hematoxylin stain that is a ferric ammonium sulfate

A

Heidenhain’s susa

92
Q

a type of iron hematoxylin stain that is for muscle striations, mitochondira, myelin, chromatin

A

Heidenhain’s susa

93
Q

a Mallory’s PTAH/Phospotungistic Acid hematoxylin

A

Tungsten hematoxylin

94
Q

For staining muscle striations alone

A

Tungsten hematoxylin

95
Q

In tungsten hematoxylin,

To ripen: stand in the _____ for several weeks or use ______ for immediate ripening

A

light

potassium

96
Q

Used for the study of spermatogenesis

A

Copper hematoxylin

97
Q

H&E staining steps elaborate

A
  1. Xylol REHYDRATE [2 changes: descending grade OH]
  2. Ammonia water submerge [Scott’s tap water]
  3. Harris/Erlich’s stain
  4. Tap water rinse
  5. 1% acid alcohol differentiator [1-2 dips]
  6. Ammonia water dip
  7. Tap water rinse
  8. Submerge OH
  9. Submerge Eosin
  10. Tap water rinse
  11. OH DEHYDRATE [4 changes: ascending grade 95,95,100, 100]
  12. Xylene- Dealcoholization-Clearing
  13. mount, label
98
Q

Removal of pigments is done after _______ and right before ______

A

rehydration

primary staining

99
Q

___ -1 stain [nuclear]
___- 2 counterstain [cytoplasmic]

A

Hematoxylin

Eosin

100
Q

bridging mordant and dye that will
intensify the color of the stain

A

Blueing step

101
Q

Reagents commonly used in staining procedure

A
  1. Acid alcohol [differentiator]
  2. Ammonia water [bluing]
  3. OG-6 [stains cytoplasm of mature superficial cells]
  4. Eosin azure [intermediate, para-basal, immature cells]
102
Q

color bearers _____
auxochromes _____

A

chromophores
auxochromes

103
Q

When attracted to the dye molecule, they serve to intensify the color of the dye. They do this by acting as electron donors to the chromophore

A

auxochromes

104
Q

the resultant complex of stain-mordant-tissue

A

Lake

105
Q

Papanicolau’s stain was developed by [who] [when]

A

George Papanicolau [1942]

106
Q

George Papanicolau published 3 formulations of this stain [3 dates]

A

1942, 1954, 1960

107
Q

Pap’s smear uses ____________staining
technique

A

multichromatic cytological

108
Q

One of the most widely used stains in
cytology, where it is used to aid pathologists in making a diagnosis

A

Pap’s smear

109
Q

Although most notable for its use in the detection of ________in the Pap test or Pap smear

A

cervical cancer

110
Q

If fluid cytology/ slides – ______

If cell block- ______

A

Pap’s smear

Routine H&E

111
Q

Pap’s smear is also used to stain _______
specimen preparations from a variety of bodily ______ and from small needle _______ and organs and tissues

A

non-gynecological spx prep

fluids
biopsies

112
Q

Pap smear stains [3]

A

Hematoxylin
Orange G-6
Eosin Azure- 50

113
Q

Pap smear procedure

A

ELEMENTS:
Fix: 95% alcohol for 1 min. Wash: water
Dip: Xylol for 5-10-30 mins
Stains:
Harris hematoxylin: 20-30 mins
OG-6: 3-5 mins
Eosin Azure-50: 3-5mins

PROCEDURE:
1-2: Fix-wash
3-5: Fix- Stain [Harris 20-30 mins]- Wash
6-8: Fix- Stain [OG-6 3-5mins]- Wash
9-11: Fix- Stain [Eosin-50 3-5mins]
12-14: Fix- Xylol dip-Mount/Label

114
Q

The classic form of the Pap’s stain involves 5 stains in 3 solutions

A

Generalized Staining Method

115
Q

In Generalized Staining Method, George Papanicolaou uses ________ in all 3 formulations of the stains he published

A

Harris hematoxylin

116
Q

Enumerate 3 formulations in the Generalized staining method

A

1st:
Hematoxylin -stains nuclei

2nd:
OG-6 + 95% ethyl alcohol + small amount of phosphotungstic acid

3rd:
Bismarck Brown Y + 95% ethyl alcohol + small amount of phosphotungstic acid + lithium carbonate
Eosin Y
Light Green SF Yellowish

117
Q

Other Formulations Include:

A

EA-36
EA-50
EA-65

118
Q

The counterstain is dissolved_______ which prevents cells from over staining which would obscure [2] especially in the case when cells are overlapping on the slide.

A

95% ethyl alcohol

nuclear detail

cell outlines

119
Q

added to adjust the PH of counterstain and helps to optimize the color intensity

A

Phosphotungistic acid

120
Q

The EA counterstain contains [2], which when in combination, cause both to precipitate out of solution, reducing the useful life of the mixture

A

bismarck brown
phosphotungstic acid

121
Q

➔ The stain should result in cells that are fairly _______ so even thicker specimens with overlapping cells can be interpreted
➔ Cell nuclei should be ______, ______in color and the ________ of the nucleus should be well defined.
➔ Cell cytoplasm stains _______ and keratin stains ______ in color

A

transparent

crisp
blue-black
chromatin patterns

blue-green
orange

122
Q

Stains the superficial epithelial squamous cells, nucleoli, cilia, and RBC

A

Eosin Y

123
Q

Stains the cytoplasm of other cells, other superficial squamous cells

A

Light Green SF Yellowish

124
Q

In Light Green SF Yellowish, superficial cells stains _______

In Light Green SF Yellowish, intermediate & parabasal cells _______

A

orange- pink

turquoise green-blue

125
Q

Used for staining hemoglobin

A

Benzidine

126
Q

has affinity to DNA/RNA

has green fluorescence _______

has red fluorescence ________

A

Acridine Orange

DNA

RNA

127
Q

Crystal Violet
For _____ in frozen sections and ______ in blood

A

amyloid

platelets

128
Q

Formed by the mixture of crystal violet, methyl violet, and dextrin

A

Gentian violet

129
Q

Stained for axis cylinders in embryos

A

Congo Red

130
Q

Congo Red is used as a 4% aqueous solution in _____ methods for staining [2]

A

Krajan

elastic tissue
myelin

131
Q

Probably the oldest of all stains

A

iodine

132
Q

Iodine– stains for [5], and a widely used for removal of [1]

A

amyloid
carotene
cellulose
starch
glycogen

mercuric fixative pigments

133
Q

Contrast stain for staining Ascaris eggs and erythrocytes, bacterial spore stain

A

Malachite green

134
Q

Demonstrate mitochondria during intravital stain

A

Janus Green

135
Q

substitute for carbol fuchsin in acid fast staning

A

Night blue

136
Q

Demonstrate neuroglia in frozen section

A

Victoria Blue

137
Q

Not real dyes, no auxochromes groups, give color to lipids simply because they are more soluble in lipid medium of the tissues than in their medium of ___% alcohol

A

Lysochromes [Oil-soluble dyes]

70%

138
Q

Examples of Oil-Soluble Dyes Used for Demonstration of Intracellular Fats [+color]

A

Sudan Black B- black
Sudan III- Orange
Sudan IV [Scharlach R]- Red

HPCT (5 decks)

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