Histopathology Staining

Staining

Process of applying dyes on the sections to see and study architectural pattern of tissues and cells

Acidic parts

Have greater affinity for basic dyes

Basic parts

Greater affinity for acidic dyes

Histological staining

Demonstration of tissue parts through direct interaction with a dye or staining solution

True

T or F:

Histological Staining involves the Coloration of the active tissue component

Histochemical staining

Studying various tissue parts through chemical reactions that will permit microscopic localization of a tissue substance.

Immunohistochemical Staining

Allows phenotypic markers to be detected under a microscope using labeled antibodies

Direct Staining

Giving color to sections using Aqueous or Aniline stains which does not require a mordant to stain.

Indirect staining

Intensification of dye action by adding mordant and accentuator

Mordant

Link or bridge between the tissue and the dye

Accentuator

Accelerates the speed of staining by increasing staining power and selectivity of the dye.

Progressive staining

The tissue is left in the stain just long enough to reach the proper end point.

Regressive staining

Tissues are overstained first to obliterate cell details, then removal of the excess stain from unwanted parts.

Metachromatic Staining

Use of specific dyes which stain substances with a color different from the stain itself.

Metallic impregnation

Using solutions of metallic salts which are reduced by the tissue

Production of black deposits on the surface of the tissue or bacteria.

Metallic impregnation

Not absorbed by a tissue

Metallic impregnation

Held physically on the surface as a precipitate or as a reduction product.

Vital staining

Staining of living cell parts, cytoplasmic phagocytosis and true vital staining.

Natural dyes

From plants and Animals

Hematoxylin

Cochineal Dyes

Orcein

Synthetic Dyes

Acidic dyes

Basic dyes

Neutral dyes

Hematoxylin

Extracted from the core of logwood

Haematoxylum campechianum

Hematoxylin is derived from what log wood?

Hematoxylin

Powerful nuclear and chromatin staining capacity.

Hematin

Active coloring agent formed after oxidation or ripening.

Hematoxylin

Used in combination with mordants like Alum and Iron

Cochineal Dye

Extracted from the female cochineal bug

Dactylopious coccus

Female cochineal bug

Cochineal Dye

a red dye used to color cotton and camelid (alpaca and llama) yarn and cloth. The dye is made from an insect that feeds on cacti in Central and South America.

Cochineal dye

Treated with alum to produce Carmine

Orcein

Vegetable dye extract from orchil lichens

Orcein

Used to stain elastic fibers (orcein)

Ehrlich's and Harris

Cole's and Mayer's

Four types of aluminum hematoxylin

Ehrlich's Hematoxylin

Ripened with sodium iodate on a regressive staining.

Harris hematoxylin

Ripened with mercuric chloride on a Regressive and progressive

widely used for routine nuclear staining.

Cole's Hematoxylin

In sequence with Celestine blue ripened with Alcoholic iodine

Mayer's hematoxylin

Ripened with Sodium Iodate

Regressive and Progressive

Gills hematoxylin

Ripened with Sodium Iodate

Progressive

Delafield’s Hematoxylin

Natural ripening

Progressive

Sodium Iodate

Ripens Ehrlich's and Mayer's

Alcoholic Iodine

Ripens Cole's

Mercuric Chloride

Ripens Harris

Weigert's Hematoxylin

Heidenhain's hematoxylin

Phosphotungstic acid Hematoxylin

Three types of Iron Hematoxylin

Weigert's hematoxylin

Uses Ferric chloride as a mordant

Used in Muscle and connective tissue fibers

Heidanhain's hematoxylin

Uses Ferric ammonium sulfate

Used for mitochondria, muscle striations, chromatin, myelin

Tungsten Hematoxyin

Mallory’s phosphotungstic Acid Hematoxylin PTAH

Tungsten Hematoxyin

Iron hematoxylin that is used for muscle striations only

Phosphotungstic acid

Natural or potassium manganate

Mordant and ripening agent used in Tungsten Hematoxyin

Eosin

Used for differentially staining connective tissues and cytoplasm, counterstain after hematoxylin or secondary stain

Eosin Y

Yellowish most commonly used Eosin

Eosin B Erythrosin B

Bluish Eosin

Eosin ES

Ethyl eosin

Blue or blue black

Nuclei

Dark blue cytoplasm

kayrosome

bright orange-red

RBCs, eosinophilic granules, keratin

purplish pink

basophilic cytoplasm, plasma cells, osteoblast

pink or light blue to dark blue

cartilage

purplish blue

Calcium and calcified bone

pink

Decalcified bone matrix, collagen, osteoid

Deep pink

Muscle fibers