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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
Q

Example of counterstain

A

Eosin - for the cytoplasm

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26
Q

Selective staining of living cell constituents, demonstrating cytoplasmic structures by phagocytosis of the dye particle (Cytoplasmic Phagocytosis)

A

Vital Staining

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27
Q

The __________ of the living cell is resistant to vital stains, and therefore is not demonstrated

A

nucleus

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28
Q

Examples of Vital Staining

A
  1. Trypan blue - vital stain for Reticuloendothelial system)
  2. Janus Green B - true vital staining of mitochondria
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29
Q

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

A

Intravital staining

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30
Q

Common dyes used in Intravital staining:

A

(LIC)
1. Lithium
2. India ink
3. Carmine

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31
Q

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

A

Supravital staining

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32
Q

Example of supravital staining

A

Neutral Red - the best vital dye

33
Q

Makes use of heavy metals which are precipitated with selectivity of certain cellular and tissue components

A

Metallic impregnation

34
Q

Has its greatest application in tissue from the CNS and for the demonstration of reticulin

A

Metallic Impregnation

35
Q

Differs from staining because it consists of an opaque black particulate precipitate

A

Metallic Impregnation

36
Q

Examples of Metallic Impregnation

A
  1. Silver Nitrate - most commonly used agent for impregnation
  2. Osmium tetroxide - used for demonstration of lipids
37
Q

these dyes are obtained from plants and animals, previously utilized for dyeing of wool and cotton

A

Natural dyes

38
Q

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

A

Hematoxylin

39
Q

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

40
Q

Mainly used for staining Elastic fibers

41
Q

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

42
Q

A plant with orange stigmas yielding a dye

43
Q

Sometimes known as “coal tar dyes”

A

Synthetic dyes

44
Q

Synthetic dyes were originally manufactured from substances that have been taken from _____

45
Q

Derived from the hydrocarbon benzene and are collectively known as Aniline dyes

A

Synthetic dyes

46
Q

“color bearers”

A

Chromophores

47
Q

Substances capable of producing visible colors

A

Chromophores

48
Q

“color increaser”

A

Auxochromes

49
Q

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

A

Auxochromes

50
Q

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

A

acid and base

51
Q

A process of selective removal of excess dye

A

Differentiation

52
Q

If the dye is a basic one, differentiation is carried out by ______

A

an acid solution

53
Q

Hematoxylin is extracted from the core of heartwood of the tree _________

A

Haematoxylon campechianum

54
Q

formed by oxidation of hematoxylin

55
Q

The complex of stain and mordant is called a

56
Q

Examples of Mordants

A

(ACIC TF)
1. Aluminum
2. Chromium
3. Iron
4. Copper
5. Tungsten/Phosphotungstic acid (PTAH)
6. Ferric salt

57
Q

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

58
Q

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

A

Ferric salt

59
Q

A red acid dye for cytoplasm differentially

60
Q

Commonly used as a background or contrast stain because it gives a leasing and useful contrast to nuclear stains such as hematoxylin

61
Q

Examples of Eosin

A
  1. Eosin Y (most commonly used)
  2. Eosin B (rarely used)
  3. Eosin S/Ethyl Eosin (rarely used)
62
Q

Hematoxylin and Eosin color reaction - Cytoplasm & proteins in edema fluid

63
Q

Hematoxylin and Eosin color reaction - Decalcified bone, Osteoid, Collagen

64
Q

Hematoxylin and Eosin color reaction - Muscle fiber

65
Q

Hematoxylin and Eosin color reaction - RBC, Eosinophil granules, Keratin

A

Bright orange to Red

66
Q

Hematoxylin and Eosin color reaction - Calcium & Calcified bone

A

Purplish blue

67
Q

Hematoxylin and Eosin color reaction - Plasma cells, Osteoblast, Basophilic cytoplasm

A

Purplish pink

68
Q

Hematoxylin and Eosin color reaction - Karyosome

69
Q

Hematoxylin and Eosin color reaction - Nucleus

A

Blue to blue-black

70
Q

Hematoxylin and Eosin color reaction - Cartilage

A

Light blue to Dark blue

71
Q

A specialized fixative used in frozen section which serves to localize antigens and hydrolytic enzymes; it is also used for preservation of lipids

A

Formol Calcium

72
Q

It is similar with H and E technique but the differentiation stage is omitted; a progressive form of staining

A

Modified H and E technique

73
Q

Papanicolaou stain makes use of 3 stains:

A
  1. Hematoxylin
  2. OG-6 (orange green)
  3. Eosin Azure
74
Q

Among the three stains used in papanicolaou stain, this stain is for staining nuclear structures

A

Hematoxylin

75
Q

Among the three stains used in papanicolaou stain, this stain is for staining cytoplasm of mature cells (e.g. Mature superficial cells)

A

OG-6 (orange green)

76
Q

Among the three stains used in papanicolaou stain, this stain is for staining cytoplasm of immature cells (e.g. Intermediate and parabasal cells)

A

Eosin azure

77
Q

Composition of Eosin Azure

A
  1. Eosin
  2. Bismarck brown
  3. Lithium carbonate
  4. PTA
  5. Light green stain (36,50,65)
78
Q

This stain omitted the Bismarck brown dye for Eosin Azure solution

A

Modified Pap’s Stain

79
Q

Sharpness of color and brilliant staining reactions are improved

A

Modified Pap’s Stain

HISTOPATH LEC STAINING (3 decks)

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