STAINING Flashcards

1
Q

Three major Groups of Staining Tissues

A
  1. Histological staining
  2. Histochemical staining
  3. Immunohistochemical staining
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2
Q

Process whereby the tissue constituents are demonstrated in sections by direct interaction with a DYE or STAINING SOLUTION , producing coloration of the active tissue substance

A

Histological staining

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3
Q

Process whereby various constituents of tissues are studied through CHEMICAL REACTIONS that will permit microscopic localization of a specific tissue substance

A

Histochemical staining

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4
Q

Combination of immunologic and histochemical techniques that allow PHENOTYPIC MARKERS to be detected and demonstrated under the microscope using a wide range of polyclonal or monoclonal, fluorescent-labeled or enzyme-labeled antibodies

A

Immunohistochemical staining

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5
Q

METHODS OF STAINING:

A
  1. Direct staining
  2. Indirect staining
  3. Progressive staining
  4. Regressive staining
  5. Metachromatic staining
  6. Metallic impregnation
  7. Negative staining
  8. Counterstaining
  9. Vital staining
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6
Q

Process of giving color to the sections by using AQUEOUS or ALCOHOLIC DYE SOLUTIONS (ex. Methylene blue)

A

Direct staining

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7
Q

Process whereby the action of the dye is intensified by ADDING ANOTHER AGENT OR A MORDANT

A

Indirect staining

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8
Q

Serves as the LINK or BRIDGE between the tissue and the dye, to make the staining reaction possible

A

Mordant

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9
Q

Examples of mordant include:

A
  • Phosphotungstic acid
  • Iron
  • Lead
  • Aluminum
  • Molybdenum
  • Copper
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10
Q

What is the mordant of Harris hematoxylin?

A

Potassium aluminum

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11
Q

Accelerates or hastens the speed of the staining power and selectivity of the dye

A

Accentuator

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12
Q

The combination of a dye and a mordant

A

Lake

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13
Q

REMEMBER!

Direct staining - DO NOT USE MORDANT

Indirect staining - USES A MORDANT

A
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14
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 9ex. Modified H&E stain)

A

Progressive staining

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15
Q

The tissue is first overstained to obliterate the cellular details, and the EXCESS STAIN IS REMOVED OR DECOLORIZED from unwanted parts of the tissue, until the desired intensity of color is obtained

A

Refressive staining

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16
Q

The most common method using the regressive staining which consists of overstaining the nuclei, followed by removal of excessive color of the tissue constituent by acid differentiation

A

ROUTINE HEMATOXYLIN AND EOSIN (H&E) STAINING

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17
Q

When using the regressive staining method, the nucleus is deliberately overstained with _______

A

Non-acidified hematoxylin

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18
Q

Selective REMOVAL OF EXCESS STAIN from the tissue during regressive staining

A

DIFFERENTIATION/DECOLORIZATION

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19
Q

Removal of UNWANTED ARTIFACTS during staining

A

WASHING-OUT

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20
Q

technique using specific dyes which differentiate particular substances by staining them with a color that is DIFFERENT FROM THE STAIN ITSELF

This is particularly employed for staining cartilage, connective tissue, epithelial mucins, mast cell granules, and amyloid
(ex. Toluidine blue: stains the mast cells purple)

A

METACHROMATIC STAINING

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21
Q

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 or bacteria; the most valuable metals for this purpose are GOLD (gold chloride) and Silver (silver nitrate)

A

Metallic impregnation

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22
Q

An acidic, anionic dye is used to change the color of the background, not the cells, causing the cells to stand out; this can be achieved by smearing the sample onto the slide and then applying NIGROSIN, a black synthetic dye or INDIA INK, an aqueous suspension of carbon particles

A

NEGATIVE STAINING

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23
Q

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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24
Q

REMEMBER!!

PROGRESSIVE vs REGRESSIVE

  • Progressive staining WITHOUT differentiator/decolorizer and washing-out; ex: MODIFIED H&E stain
  • Regressive staining WITH differentiator/decolorizer and washing-out; ex: ROUTINE H&E stain
A
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25
Q

REMEMBER!!
WASHING OUT

Fixation - washing out is removing excess fixative

Staining - washing out is removing unwanted artifiacts

A
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26
Q

NUCLERA COUNTERSTAINS

RED:
BLUE:

A

NUCLERA COUNTERSTAINS

RED: “SoNCH”
- Safranin O
- Neutral red
- Carmine
- Hematoxylin

BLUE: “BLUE”
- Methylene BLUE
- Toluidine BLUE
- Celestine BLUE

27
Q

CYTOPLASMIC COUNTERSTAINS

RED:
YELLOW:
GREEN:

A

CYTOPLASMIC COUNTERSTAINS

RED: “PEER”
- Phloxine B
- Eosin Y
- Eosin B
- R - RED

YELLOW: “PORY”
- Picric acid
- Orange G
- Rose Bengal
- Y - YELLOW

GREEN: “GREEN”
- Light GREEN SF
- Lissamine GREEN

28
Q

The selective staining for LIVING CELL constituents

A

VITAL STAINING

29
Q

Types of Vital stains:

A

INTRAVITAL STAINING
SUPRAVITAL STAINING

30
Q

type of vital stain done by INJECTING THE DYE:

A

Intravital staining

31
Q

Examples of INTRAVITAL STAIN:

A

Intravital stains: “LIC”

  • Lithium
  • India ink
  • Carmine
32
Q

Type of vital stain done after REMOVAL FROM THE BODY

A

Supravital stain

33
Q

Examples of SUPRAVITAL STAINS:

A

Supravital stains:
“TTT ni JANUS kulay RED”

  • Trypan blue
  • Thionine
  • Toluidine blue
  • Nile blue
  • JANUS green
  • Neutral RED
34
Q

REMEMBER!!

JANUS GREEN - for MITOCHONDRIA

Neutral RED - for BEST VITAL DYE

Mordant of Harris hematoxylin: POTASSIUM ALUMINUM

A
35
Q

Examples of natural dyes

A
  1. Hematoxylin
  2. Orcein
  3. Cochineal Dyes
  4. Saffron
36
Q

Derived by extraction from the core of the heartwood of a MEXICAN LOGWOOD TREE known as Haematoxylon campechianum

A

HEMATOXYLIN

37
Q

The active dye or coloring agent in hematoxylin solution is _______ formed by _________ of hematoxylin in a process known as ___________

A

The active dye or coloring agent in hematoxylin solution is HEMATEIN formed by OXIDATION of hematoxylin in a process known as RIPENING

38
Q

Exposing the substance to AIR or SUNLIGHT and usually a slow process, sometimes taking as long as 3 to 4 months

A

NATURAL RIPENING

39
Q

Chemical oxidizing agents convert the hematoxylin to hematein rapidly, so these solutions are ready for use immediately after preparation

A

ARTIFICIAL/CHEMICAL RIPENING

40
Q

Arificial ripening agents include:

A
  • Sodium iodate
  • Hydrogen peroxide
  • Iodine
  • Mercuric oxide
  • Potassium permanganate

mnemonic: SHIMP

41
Q

ph can affect the oxidation or ripening process:

BASIC pH:
ACID pH:

A

ph can affect the oxidation or ripening process:

BASIC pH: MORE RAPID OXIDIZING PROCESS
ACID pH: SLOWER OXIDIZING PROCESS

42
Q

FOUR TYOPES OF HEMATOXYLIN

A
  1. ALUMINIM (ALUM) HEMATOXYLIN
  2. IRON HEMATOXYLIN
  3. TUNGSTEN HEMATOXYLIN
  4. COPPER HEMATOXYLIN
43
Q

Most frequently used hematoxylin in the H&E stain

A

Aluminum (Alum) hematoxylin

44
Q

Aluminum (Alu) hematoxylin dyes:

A
  1. Ehrlich’s hematoxylin
  2. Harris hematoxylin
45
Q

Ehrlich’s hematoxylin

USE:
RIPENING AGENT:
MORDANT:

A

Ehrlich’s hematoxylin

USE: Regressive staining
RIPENING AGENT: SODIUM IODATE
MORDANT: POTASSIUM ALUM

46
Q

Harris hematoxylin

USE:
RIPENING AGENT:
MORDANT:

A

Harris hematoxylin

USE: For exfoliative cytology
RIPENING AGENT: MERCURIC OXIDE
MORDANT: POTASSIUM ALUM

47
Q

Hematoxylin used only for REGRESSIGVE STAINING

A

IRON HEMATOXYLIN

48
Q

Types of Iron hematoxylin:

A
  1. Weigert’s hematoxylin
  2. Heidenhain’s hematoxylin
49
Q

Weigert’s hematoxylin

USE:
RIPENING AGENT:
MORDANT:

A

Weigert’s hematoxylin

USE: Standard iron hematoxylin
RIPENING AGENT: FERRIC CHLORIDE
MORDANT: FERRIC CHLORIDE

50
Q

Heidenhain’s hematoxylin

USE:
RIPENING AGENT:
MORDANT:

A

Heidenhain’s hematoxylin

USE: For nuclear and cytoplasmic inclusions
RIPENING AGENT: Ferric ammonium sulfate
MORDANT: Ferric ammonium sulfate

***Heidenhain’s hematoxylin - for tumor biopsy of the skin

51
Q

Hematoxylin stain used only for PROGRESSIVE STAINING

A

TUNGSTEN HEMATOXYLIN

52
Q

Hematoxylin stain utilized for the study of SPERMATOGENESIS:

A

COPPER HEMATOXYLIN

53
Q

Vegetable dye extracted from LICHENS (colorless) but when treated with ammonia and exposed to air, produces blue or violet colors

A

ORCIEN

54
Q

Old histologic dye derived from an extract from the female COCHINEAL BUG 9Coccus cacti)’

A

COCHINEAL DYES

55
Q

If Cochineal dye is treated with alum, it will produce a dye called _______ which is used for glycogen demonstration:

A

CARMINE DYE

56
Q

Derived from the dried stigmata of FLOWER, Crocus Sativus (known popularly as Saffron crocus)

A

SAFFRON

57
Q

Synthetic dyes are sloknown as:

A

COAL TAR DYES

58
Q

Synthetic dyes are derived from _____ and are collectively known as ANILINE DYES

A

Synthetic dyes are derived from BENZENE and are collectively known as ANILINE DYES

59
Q

EXAMPLES OF SYNTHETIC DYES:

A
  1. Chromopore
  2. Auxochromes
60
Q

Substances that are capable of producing VISIBLE colors; responsible for the COLORING PROPERTY OF THE DYE

A

CHROMOPHORE

61
Q

Impart color to the tissue that is NOT PERMANENT and can be EASILY REMOVED

A

CHROMOGENS

62
Q

Substances added to chromogen that alters its property enabling it to RETAIN ITS COLOR in the tissues; responsible for the DYEING PROPERTY OF THE DYE

A

AUXOCHROMES

63
Q

REMEMBER!!!

A DYE should consist of an AUXOCHROME and a CHROMOPHORE group attached to hydrocarbon benzene ring

A