HPCT > Staining > Flashcards

Staining Flashcards

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1
Q

*Renders the different tissue constituents more visible.

A

Staining

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2
Q

*Promoting easier optical differentiation & identification
of the cell & tissue components.

A

staining

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3
Q

Major Groups of Staining of Tissues

A

*Histological Staining
*Histochemical Staining (Histochemistry)
* Immunohistochemical Staining

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4
Q

*Tissue constituents are demonstrated in sections by direct
interaction with a dye or staining solution, producing
coloration of the active tissue component.

1
2
3

A

Histological Staining
*Microanatomic stains
* bacterial stains
* specific tissue stains (muscles, connective tissue and neurologic stains

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5
Q
  • thru chemical reactions that will permit microscopic
    localization of a specific tissue substance.
A

histochemical Staining (Histochemistry)

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6
Q

*Perl’s prussian blue reaction for __

A

Histochemical
- hemoglobin

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7
Q

*Periodic Acid Schiff staining for

A

Histochemical
-carbohydrates

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8
Q

*In ___,
*the active reagent serves as a substrate upon which the
enzymes act,
*the final opacity or coloration produced from the
substrate rather than the tissue

A

Histochemical Staining (Histochemistry)
- Enzyme histochemistry

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9
Q
A
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10
Q

*A combination of immunologic and histochemical techniques that allow phenotypic markers to be detected
and demonstrated under the microscope.
1
2

A

Immunohistochemical Staining
*flourescent labeled
* enzyme-labeled antibodies

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11
Q

*The process of giving color to the sections by using aqueous or alcoholic dye solutions
1
2

A

Direct Staining
(methylene blue, eosin)

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12
Q

*The process whereby the action of the dye is intensified by adding another agent or mordant, which serves as a link or bridge between the tissue and the dye, to make the
staining reaction possible.

A

Indirect Staining

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13
Q

*Example of mordants:

A
  • potassium alum with hematoxylin in Ehrlich’s hematoxylin,
  • iron in Weigert’s hematoxylin
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14
Q

*Does not participate in the staining reaction, but it accelerates
or hastens the speed of the staining reaction by increasing the
staining power and selectivity of the dye.

A

Indirect Staining
* Accentuator

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15
Q

*Examples of accentuators:

A
  • potassium hydroxide in Loeffler’s methylene blue and
  • phenol in carbol thionine and carbol fuchsin
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16
Q

*The process by which tissue elements are stained in a
definite sequence

A

Progressive Staining

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17
Q
  • the staining solution is applied for specific periods of
    time or until the desired intensity of coloring of the
    different tissue elements is attained.
A

Progressive Staining

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18
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

Regressive Staining

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19
Q

*Is the selective removal of excess stain from the tissue
during regressive staining.

A

Differentiation (Decolorization)

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20
Q

*Usually done by washing the section in simple solution (water
or alcohol), or by the use of acids or oxidizing agents.

A

Differentiation (Decolorization)

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21
Q

– tissue stains in color shades that are
similar to the color of the dye itself.

A

orthochromatic

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22
Q

entails the use of
specific dyes which differentiate particular substances by
staining them with a color that is different from that of
the stain itself (__)

A

Metachromatic staining technique
- metachromasia

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23
Q

Metachromatic Staining
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A
  • for staining cartilage,
  • connective tissues,
  • epithelial mucins,
    *mast cell granules,
  • amyloid
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24
Q
A
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25
Metachromatic Dyes
1. Methyl violet or crystal violet 2. Cresyl blue (for reticulocytes) 3. Safranin 4. Bismarck brown 5. Basic fuchsin 1.Methylene blue 2. Thionine 3. Toluidine blue 4. Azure A, B, C
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The application of a different color or stain to provide contrast and background to the staining of the structural components to be demonstrated.
Counterstaining
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CYTOPLASMIC
red yellow green
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* Red cytoplasmic counterstaining
* Eosin Y & B * Phloxine B
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* Yellow cytoplasmic counterstaining
* Picric acid * Orange G * Rose Bengal
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* Green cytoplasmic counterstaining
* Light green SF * Lissamine green
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NUCLEAR
red blue
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* Red nuclear counterstaining
* Neutral red * Safranin O * Carmine * Hematoxylin
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* Blue nuclear counterstaining
* Methylene blue * Toluidine blue * Celestine blue
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*A process where specific tissue elements are demonstrated, not by stains, but by colorless solutions of metallic salts.
Metallic Impregnation
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- forming black deposits seen under the microscope
Ammoniacal silver
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is different from a stain. It is not absorbed by the tissue, held physically on the surface as a precipitate. It is a reduction product in certain tissue components
metallic impregnating agent
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Metallic Impregnation *Most valuable metal:
* gold (gold chloride) * silver (silver nitrate
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*Selective staining of living cell constituents, * demonstrating cytoplasmic structures by phagocytosis of the dye particle. * the staining of mitochondria by __
Vital Staining - Janus green
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*Done by injecting the dye into any part of the animal body (intravenous, intraperitoneal, or subcutaneous)
Intravital Staining
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* producing specific coloration of certain cells, particularly those of the reticulo-endothelial system *Common dyes used: 1 2 3
Intravital Staining * lithium, carmine and India ink
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* Used to stain living cells immediately after removal from the living body.
Supravital Staining
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Supravital Staining Common dyes used:
* Neutral red – BEST VITAL DYE * Janus green - MITOCHONDRIA * Trypan blue - * Nile blue * Thionine * Toluidine blue
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*Most common method utilized for microanatomical studies, using regressive staining
H & E Staining Technique
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* 4 Staining Method for Frozen section
* Hematoxylin-Eosin method * Thionine * Polychrome Methylene Blue * Alcoholic Pinacyanol method >Supravital staining
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2 Categories of Dyes
1. Natural dyes: 2. Synthetic dyes:
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Obtained from plants or animals
Natural dyes:
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Aniline or Coal Tar dyes
Synthetic dyes:
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Natural Dyes
*Hematoxylin (hematoxylin campechianum) * Cochineal dyes (female cochineal bug) *Orcein *Saffron
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* tx. With alum = ___ * Combine with picric acid (___) * _ - Best carmine stain
Cochineal dyes (female cochineal bug) - carmine - picrocarmine - Aluminum chloride
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* Vegetable dye * Used for staining elastic fibers
Orcein
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*A natural dye *Most valuable staining reagent used by the cytologist *Active coloring agent is __ *Most commonly used for routine histologic studies
Hematoxylin ( Hematoxylin campechianum) - hematin
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*Is a weak anionic dye. *Has little affinity for tissues but become a strong nuclear dye when combined with a metallic mordant
Hematein
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Harris Hematoxylin
M Ammonium Aluminum Sulfate O Sodium Iodate
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Mayer Hematoxylin and Ehrlich Hematoxylin
M Potassium Aluminum Sulfate or Ammonium Aluminum Sulfate O Sodium Iodate
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Gill Hematoxylin
M Aluminum Sulfate, Al(SO4)3.18H20 O Sodium Iodate
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are substances with definite atomic groupings and are capable of producing visible colors
Chromophores
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- consist of chromophore (coloring property) and auxochrome (dyeing property)
Dye
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Acid Dyes
Picric acid – fix, differentiate, stain tissue
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Basic Dyes
Methylene blue – indicator , dye
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Capable of staining cytoplasm and nucleus
Neutral Dyes a) Romanowsky b) Giemsa’s c) Irishman’s
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* *where the coloring substances are found in the acid component and sodium is the agent used to remove the base radical.
Acid Dyes:
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* * where the coloring substance is found in the basic component, * the acid radical usually taken by sulfuric or hydrochloric acid.
Basic Dyes:
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* *When an acid dye is combined with a basic dye * *As it contains both coloured radicals, it gives * different colours to cytoplasm and nucleus
Neutral Dyes:
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*Routinely used in histopathology as a counterstain; used for differentially staining connective tissues and cytoplasm
Eosin
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*Commonly used as a background stain. EOSIN
* Yellowish (Eosin Y) – most commonly used * Bluish (Eosin B, Erythrosin B) – deeper red color * Ethyl eosin – eosin s, eosin alcohol-soluble
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*Mixture of picric acid and acid fuchsin for the demonstration of connective tissues
Van Gieson Stain
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*Basic acridine fluorochrome which permits discrimination between dead and living cells 1 2
Acridine Orange *Green fluorescence – DNA * Red fluorescence - RNA
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*For the demonstration of calcium salts deposits and possible sites of phosphatases activities
Acridine Red 3B
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*Stains acid mucopolysaccharides by forming salt linkages *Excellent stain because it is simple and produces a striking blue color * It is more specific for connective tissues and epithelial mucin
Alcian Blue
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*Cytoplasmic stain used for counterstaining epithelial sections
Aniline Blue
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- Used for staining hemoglobin
Benzidine
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Contrast stain for Gram’s technique in acid fast and Papanicolau method, - Stains diptheria organisms
Bismarck Brown
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, - Stains diptheria organisms
Bismarck Brown
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- Best known as an indicator, but may be utilized as a stain for axis cylinders in embryos
Congo Red
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- Is a nuclear or chromatin stain used for staining amyloid in frozen sections and platelets in blood
Crystal Violet
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– formed by the mixture of crystal violet, methyl violet and dextrin
Gentian violet
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-used for staining blood to differentiate leukocytes
Giemsa Stain
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- oldest of all stains, originally used for microscopic study of starch granules *Stains__,__,__,__,__
Iodine amyloid, cellulose, starch, carotenes and glycogeN
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- demonstrating mitochondria during intravital staining
Janus Green B
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- weakly basic dye used as a contrast stain for staining ascaris eggs and erythrocytes and bacterial spore stain, Also a decolorizer
*Malachite Green
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- a common basic nuclear stain employed with eosin to provide marked differentiation of various structures in the tissue * Contains azures or methylene violet
*Methylene Blue
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* Excellent stain for elastic fibers * Recommended for dermatological studies due to its ability to demonstrate the finest and most delicate fibers in the skin
Orcein
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* It is used with osmic acid to fix and stain blood and glandular tissues
*Rhodamine B
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* Nuclear stain for fixed tissues * Recommended for staining of Nissl granules or chromatophilic bodies
* Toluidine Blue
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* For the demonstration of neuroglia in frozen sections
*Victoria Blue
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