STAINING Flashcards by Eldren Joi Malsi

Render the different tissue constituents more visible, thru variation in colors, promoting easier optical differentiation and identification of the cell and tissue components

STAINING

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Natural dye derived from the heartwood of a Mexican tree, Hematoxylin campechianum

Hematoxylin

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Active coloring agent of hematoxylin

Hematein

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Hematoxylin used for progressive staining

Alum Hematoxylin

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Hematoxylin used for differential or regressive staining

Iron Hematoxylin

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Hematoxylin utilized for the study of spermatogenesis

Copper Hematoxylln

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Hematoxylin –[o]→ Hematin; karyosome: dark blue, nucleus: blue; cytoplasm: pink

RIPENING

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Derived from an extract of the female Cochineal bug (Coccus cacti) –[alum]→ Carmine

Cochineal Dyes

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Vegetable dye extracted from Lichens

Orcein

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Elastic fiber, dermatological studies

Orcein

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Color of Orcein + Ammonia and exposed to air

Blue or Violet color

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Derived from benzene

Synthetic (Artificial) Dyes

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Synthetic (Artificial) Dyes is also known as

Aniline dyes

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Consists of a chromophore and an auxochrome attached to a hydrocarbon benzene ring

Dye

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Substances capable of producing visible colors

Chromophores

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Benzene compounds which contain chromophores are known as

Chromogens

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Color imparted to the tissue by chromophores is permanent. True or False?

False; Not permanent

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Substances with the property of forming salts with another compound, and ultimately retaining its color

Auxochrome

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Coloring substance is found in the acid component

Acid Dyes

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Coloring substance is found in the basic component

Basic Dyes

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Formed by combining aq. solutions of acid and basic dyes, capable of staining cytoplasm and nucleus simultaneously and differentially, usually soluble in alcohol but insoluble in water

Neutral Dyes

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Using potassium alum; for regressive staining, MPS substances (e.g., cartilage, cement lines of bones) are stained intensely blue

Ehrlich’s hematoxylin

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Using ammonium or potassium alum; for routine nuclear staining, in exfoliative cytology, and for staining sex chromosomes

Harris hematoxylin

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Using ammonium alum; used in Celestine Blue Hemalum Method of nuclear staining

Cole’s hematoxylin
Mayer’s hematoxylin

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Using ferric ammonium chloride (iron alum); for demonstrating muscle fibers and connective tissues

Weigert's hematoxylin

Using ferric ammonium sulfate (iron alum); for demonstrating nuclear and cytoplasmic inclusions e.g., chromatin, chromosomes, nucleoli, centrosomes, and mitochondria

Heidenhain's hematoxylin

Used as counterstain after Hematoxylin and before Methylene Blue

Eosin

2 shades of Eosin

Eosin B—bluish Eosin Y—most commonly used, yellowish

Nervous tissue, diagnosis of diphtheria

Methylene Blue

Metachromatic dye formed when Methylene Blue is heated; WBCs

Methylene Violet

Nissl’s granules or chromophiIic bodies

Toluidine Blue

Amyloid in frozen sections and platelets in blood

Crystal Violet

Counterstaining of epithelial sections

Aniline Blue

Acid-fast organisms, mitochondria

Basic Fuchsin

Masson Stain

Acid Fuchsin

Blood to differentiate WBC’s

Giemsa Stain

Staining of fixed sections

Celestine Blue

Stains chromatin green in the presence of an acid

Methyl Green

Contrast stain for Ascaris eggs and RBCs, and as a bacteriaI spore stain

Malachite Green

Contrast stain for Gram's, acid-fast, and Papanicolau methods, and for staining Diphtheria organisms

Bismarck Brown

Intravital staining

Prussian Blue

Connective tissue

Picric Acid

Carmine stains

Best Carmine Solution: glycogen Mucicarmine: mucin Picrocarmine: neuropathoIogicaI studies Azocarmine: connective tissues

Acidic subtances

Mayer’s Carmalum Solution

Amyloid, cellulose, starch, carotenes, and glycogen

Iodine

Elastic tissues, amyloid, and myelin

Congo Red

Observing cell granules and vacuoles of phagocytic cells

Neutral Red

Acid MPS

Alcian Blue

Mitochondria during intravital staining

Janus Green B

Neuroglia in frozen sections

Victoria Blue

Substitute for carbol-fuchsin in acid-fast staining

Night Blue

Demonstrate deposits of calcium salts and possible sites of phosphatase activities

Acridine Red 3b

Discrimination between dead and living cells

Acridine Orange

Color of RNA using Acridine Orange

Red

Color of DNA using Acridine Orange

Green

Hemoglobin

Benzidine

Phospholipids

Sudan Black B and Oil Red-O

Blood and glandular tissues

Rhodamine B

Triglycerides and Neutral lipids (deep red)

Sudan IV /Scharlach R

Fats (orange)

Sudan III

Metallic impregnation

Gold Sublimate Solution

Fats (stains black)

Osmium Tetroxide

Spirochetes, reticulum, and other fibers

Silver Nitrate

Demonstration of connective tissue; SIMPLEST method of differential staining of collagen

Van Gieson

Histochemical stain. Demonstration of carbohydrates (GLYCOGEN)

Periodic Acid Schiff (PAS)

Calcium: BLACK

Von Kossa Silver Nitrate

DNA

Feulgen’s stain

Phosphotungstic Acid Hematoxylin; muscles and bones; Astrocytes

Mallory’s (PTAH)

(Gomori). Differential staining of pancreatic islets of Langerhan

Aldehyde Fuschin Stain

Muscle

Lissamine Fast Red Tartrazine Method

Warthin-Starry. Spirochetes

Levaditi’s method

Melanin (Silver Modification): BLACK

Masson Fontana Technique

Diagnostic for Bile pigments

Gmelin’s Test

Hemosiderin (iron-containing pigment of hemoglobin)

Perl’s Prussian Blue

Neurons, Axons, Neurofibrils

Bielschowsky’s Technique

Normal Myelin Sheaths

Weigert-Pal Technique

Copper

Lindquist’s Modified Rhodamine

Bacteria

Gram-Twort

Helicobacter

Cresyl Violet Acetate

Bacteria, Nocardia, Actinomyces

Brown and Brenn

Legionella pneumophilia

Dieterle

Hepatitis B Surface antigen

Orcein method

Fungi

Grocott Methamine Silver

Reticular Fibers

Gordon and Sweet’s method

CHIEF SOLVENTS USED FOR STAINS

DISTILLED WATER ALCOHOL—ethyl alcohol, methyl alcohol ANILINE WATER PHENOL—used in 0.5 to 5% aq. solution

Process by which sections are stained with simple aqueous or alcoholic solutions of the dye

Direct Staining

Substances which combine with the tissue and the stain forming a bridge or link to make staining reaction possible

Mordant

Substances that accelerate or hasten the speed of the staining reaction by increasing the staining power and selectivity of the dye

Accentuator

Accentuator is essential to the chemical union of the tissue and the dye. True or False

False; Not essential

Accentuator does not participate in the staining reaction. True or False?

True

Process whereby tissue elements are stained by more than one stain in a definite sequence, and once the dye is taken up by the tissue, it is not decolorized

Progressive Staining

Process whereby tissue is first overstained to obliterate the cellular details, and the excess stain is decolorized from unwanted parts of the tissue

Regressive Staining

SeIective removal of excess stain from the tissue during regressive staining

Differentiation (Decolorization)

Process which differentiate tissue components by staining them with a color that is different from that of the stain itself (metachromasia); tissue components combine with these dyes to form a different color from the surrounding tissue

Metachromatic Staining

Application of a different stain to provide contrast and background to the staining of the structural components to be demonstrated

Counterstaining

Used to demonstrate the general relationship of tissues and cells with general differentiation of nucleus and cytoplasm

Microanatomical Staining

Demonstrates minute specific structures found in the cytoplasm and nucleus without necessarily differentiating tissue structures in general

Cytoplasmic staining

Demonstrates bacterial morphology; it is the background that is stained and not the organism;

Negative staining

Example of negative stain

India Ink

Selective staining of living cell constituents, demonstrating cytoplasmic structures by phagocytosis of the dye particle

Vital Staining

Done by injecting the dye into any part of the animal body, producing specific coloration of certain cells

Intravital Staining

Process where 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

Metallic Impregnation

Staining living cells immediately after removal from the living body

Supravital Staining

Paraffin wax is poorly permeable to most staining solutions and should be removed from the section prior to staining. True or False?

True

If aqueous stain is to be used, alcohol is finally replaced with water, before actual staining of section is performed

“SECTIONS TO WATER“

If alcoholic stain is to be used, there is no more need to replaced the alcohol with water after deparaffinization with xylene, section is subjected to decreasing grades of alcohol

"SECTIONS TO ALCOHOL”

Tissue constituents are demonstrated in sections by direct interaction with a dye, producing coloration of the active tissue componen

Histological Staining

ChemicaI constituents of tissues are demonstrated thru chemical reactions that permit microscopic localization of a specific tissue substance

Histochemical Staining

Most common method utilized for microanatomical study of tissues

H&E Staining

Recommended for connective tissues, fixed with osmium tetroxide

Phosphotungstic acid

Best stain for electron microscopy

Uranyl acetate

May be used as primary stain or as a secondary stain, following PTA or uranyl acetate

Lead