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HISTOPATH LEC STAINING > METHODS OF STAINING > Flashcards

METHODS OF STAINING Flashcards

1
Q

It is the basis of histochemistry; it is accomplished by controlled, specific chemical reactions designed to give a final color(staining) ate the site/location of the structure of the substances in thee cells or tissues

A

Specific Staining

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2
Q

Examples of Specific Staining

A
  1. Perl’s Prussian Blue reaction - hemosiderin
  2. Periodic Acid Schiff technique - polysaccharide
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3
Q

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

A

Simple Staining/Direct Staining

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4
Q

Example of Simple/Direct Staining

A

Methylene blue and Eosin

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5
Q

The action of the dye is intensified by some other agents

A

Indirect staining

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6
Q

Substance which when taken up by tissue, helps make the in return serving as a link or bridge to make the staining reaction possible

A

Mordant

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7
Q

Combines with a dye forming a colored “lake” which combines with tissue to form an insoluble “tissue-mordant-dye complex”

A

Mordant

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8
Q

An integral part of the staining reaction itself, without which, no staining could possible occur

A

Mordant

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9
Q

Examples of Mordant

A
  1. Potassium Alum with Hematoxylin in Ehrlich’s hematoxylin
  2. Iron in Weigert’s hematoxylin
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10
Q

Chemical substances that does not participate but merely increases or heightens the color intensity, selectively and crispness of the stain

A

Accentuator

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11
Q

Differ from mordants in that they do not bind or link the tissue to the dye

A

Accentuator

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12
Q

Examples of Accentuator

A
  1. Potassium hydroxide (KOH) in Loeffler’s Methylene blue
  2. Phenol in Carbol thionine and Carbol fuchsin
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13
Q

Staining is continued in a definite sequence until the desired intensity of coloring of the different tissue elements is attained

A

Progressive staining

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14
Q

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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15
Q

Example of Progressive staining

A

Any stain is possible as long as no differentiation is done

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16
Q

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

A

Regressive staining

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17
Q

What are the two principles of Regressive staining?

A
  1. Overstaining
  2. Washing/Differentiation/Decolorization
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18
Q

Examples of Regressive staining

A
  1. Acid Fast Stain
  2. Gram staining
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19
Q

Metachromatic staining is also known as

A

Metachromasia

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20
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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21
Q

Metachromatic staining is particularly employed for staining of:

A

(CECAM)
1. Cartilage
2. Epithelial mucin
3. Connective tissue
4. Amyloid
5. Mast cell granules

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22
Q

basic dye belonging to Thizine and Triphenylmethane groups

A

Metachromatic dye

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23
Q

Example of Metachromatic stain

A

Cresyl blue for reticulocytes

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24
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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25
Example of counterstain
Eosin - for the cytoplasm
26
Selective staining of living cell constituents, demonstrating cytoplasmic structures by phagocytosis of the dye particle (Cytoplasmic Phagocytosis)
Vital Staining
27
The __________ of the living cell is resistant to vital stains, and therefore is not demonstrated
nucleus
28
Examples of Vital Staining
1. Trypan blue - vital stain for Reticuloendothelial system) 2. Janus Green B - true vital staining of mitochondria
29
It is 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
Intravital staining
30
Common dyes used in Intravital staining:
(LIC) 1. Lithium 2. India ink 3. Carmine
31
Used to stain living cells immediately after removal from the living body
Supravital staining
32
Example of supravital staining
Neutral Red - the best vital dye
33
Makes use of heavy metals which are precipitated with selectivity of certain cellular and tissue components
Metallic impregnation
34
Has its greatest application in tissue from the CNS and for the demonstration of reticulin
Metallic Impregnation
35
Differs from staining because it consists of an opaque black particulate precipitate
Metallic Impregnation
36
Examples of Metallic Impregnation
1. Silver Nitrate - most commonly used agent for impregnation 2. Osmium tetroxide - used for demonstration of lipids
37
these dyes are obtained from plants and animals, previously utilized for dyeing of wool and cotton
Natural dyes
38
The most important and the most commonly used for routine histologic studies
Hematoxylin
39
A vegetable dye extracted from lichens which are normally colorless, but when treated with ammonia and expose to air, produce a blue or violet colors
Orcein
40
Mainly used for staining Elastic fibers
Orcein
41
An old histologic dye extracted from the female cochineal bug (coccus cacti)
Cochineal
42
A plant with orange stigmas yielding a dye
Saffron
43
Sometimes known as "coal tar dyes"
Synthetic dyes
44
Synthetic dyes were originally manufactured from substances that have been taken from _____
coal tar
45
Derived from the hydrocarbon benzene and are collectively known as Aniline dyes
Synthetic dyes
46
"color bearers"
Chromophores
47
Substances capable of producing visible colors
Chromophores
48
"color increaser"
Auxochromes
49
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
Auxochromes
50
For a chromogen to be a dye, it must be composed of an ____ and a ____, and therefore have salt-forming properties, ultimately retaining its color
acid and base
51
A process of selective removal of excess dye
Differentiation
52
If the dye is a basic one, differentiation is carried out by ______
an acid solution
53
Hematoxylin is extracted from the core of heartwood of the tree _________
Haematoxylon campechianum
54
formed by oxidation of hematoxylin
Ripening
55
The complex of stain and mordant is called a
Lake
56
Examples of Mordants
(ACIC TF) 1. Aluminum 2. Chromium 3. Iron 4. Copper 5. Tungsten/Phosphotungstic acid (PTAH) 6. Ferric salt
57
most commonly used mordant; gives a blue lake appearance; increases the selectivity for nuclei, esp if acid is added or is used as a differentiating agent
Aluminum
58
causes an intense blue-black appearance when used as a mordant
Ferric salt
59
A red acid dye for cytoplasm differentially
Eosin
60
Commonly used as a background or contrast stain because it gives a leasing and useful contrast to nuclear stains such as hematoxylin
Eosin
61
Examples of Eosin
1. Eosin Y (most commonly used) 2. Eosin B (rarely used) 3. Eosin S/Ethyl Eosin (rarely used)
62
Hematoxylin and Eosin color reaction - Cytoplasm & proteins in edema fluid
Pale pink
63
Hematoxylin and Eosin color reaction - Decalcified bone, Osteoid, Collagen
Pink
64
Hematoxylin and Eosin color reaction - Muscle fiber
Deep pink
65
Hematoxylin and Eosin color reaction - RBC, Eosinophil granules, Keratin
Bright orange to Red
66
Hematoxylin and Eosin color reaction - Calcium & Calcified bone
Purplish blue
67
Hematoxylin and Eosin color reaction - Plasma cells, Osteoblast, Basophilic cytoplasm
Purplish pink
68
Hematoxylin and Eosin color reaction - Karyosome
Dark blue
69
Hematoxylin and Eosin color reaction - Nucleus
Blue to blue-black
70
Hematoxylin and Eosin color reaction - Cartilage
Light blue to Dark blue
71
A specialized fixative used in frozen section which serves to localize antigens and hydrolytic enzymes; it is also used for preservation of lipids
Formol Calcium
72
It is similar with H and E technique but the differentiation stage is omitted; a progressive form of staining
Modified H and E technique
73
Papanicolaou stain makes use of 3 stains:
1. Hematoxylin 2. OG-6 (orange green) 3. Eosin Azure
74
Among the three stains used in papanicolaou stain, this stain is for staining nuclear structures
Hematoxylin
75
Among the three stains used in papanicolaou stain, this stain is for staining cytoplasm of mature cells (e.g. Mature superficial cells)
OG-6 (orange green)
76
Among the three stains used in papanicolaou stain, this stain is for staining cytoplasm of immature cells (e.g. Intermediate and parabasal cells)
Eosin azure
77
Composition of Eosin Azure
1. Eosin 2. Bismarck brown 3. Lithium carbonate 4. PTA 5. Light green stain (36,50,65)
78
This stain omitted the Bismarck brown dye for Eosin Azure solution
Modified Pap's Stain
79
Sharpness of color and brilliant staining reactions are improved
Modified Pap's Stain

HISTOPATH LEC STAINING (3 decks)

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