STAINING PT. 2

Natural Dyes

Derived from plants and animals

Cochineal Dyes, orcein, saffron, Hematoxylin

Examples of Natural Dyes

Hematoxylin

Most common

Not a stain

Extracted from tree and ripened

Hematoxylin

Formed from the oxidation of hematoxylin either by exposing it to sunlight or by adding oxidizing agents

Hematin

active coloring agent of Hematoxylin

Hydrogen peroxide, Mercuric peroxide, Sodium iodate, Sodium perborate, Potassium permanganate

Oxidizing agents of Hematoxylin

Ripened hematoxylin

has a poor staining quality and must be combined with mordant

alum, iron, chromium and copper salts

Combines with ripened hematoxylin

Alum Hematoxylin

Uses as potassium alum as mordant

Ripened hematoxylin + potassium alum

Components of Alum Hematoxylin

Ripening agent of Erhlich’s hematoxylin

Sodium iodate

Harris hematoxylin

To demonstrate sex chromosomes

Used in exfoliative cytology

Used in Papanicolaou staining

Ripening agent of Harris hematoxylin

Mercuric chloride

Ripening agent of Cole’s Hematoxylin

Alcoholic iodine

Rioening agent of Mayer’s hematoxylin

Sodium iodate

Mayer’s Hematoxylin

Used in immunohistochemistry

Used in Celestine blue hemalume method

Delafield’s

Can impart purple-red color

Smells like wine

Gills’

Used for staining mucin in goblet cells (found in columnar cells)

Ripening agent of Gills

Sodium iodate

Ripening agent of Carazzi’s

Potassium iodate

Iron Hematoxylin

In general for photomicrography

Imparts black color

Weigert’s Hematoxylin

Ferric ammonium chloride: ripening agent is also the mordant

For muscle fibers and CT

Heidenhain’s hematoxylin

Ferric ammonium sulfate: both the ripening agent and the mordant

For nuclei & cytoplasmic inclusion i.e., mitochondria

Loyez hematoxylin

Frozen sections

Verhoeff’s hematoxylin

Elastic fibers

Regaud’s Hematoxylin

For coloring mitochondria

Modification of iron hematoxylin

Copper Hematoxylin

To demonstrate spermatogenesis

Tungsten Hematoxylin

Phosphotungstic acid: mordant

Lead Hematoxylin

Demo of granules of endocrine cells of alimentary tract

Molybdenum Hematoxylin

Demo of collagen, reticulin and argentaffin cells

Cochineal dyes

Extracted from bug treated with alum to produce carmine (dye)

Picrocarmine

w/ picric acid

for neuropathological studies

Aluminum chloride

Best carmine for glycogen

Orcein

Vegetable dye extracted from lichens

Used for staining elastic fibers

Litmus

Vegetable dye extracted from lichens

Not used as cytological stain but as a pH indicator because of poor staining property

Synthetic Dyes

A.k.a. Coaltar dyes or Aniline dyes

Chromophore & Auxochrome

Components of Synthetic dye

Chromophore

For coloring property

Property of imparting color

Auxochrome

For dyeing property

Property of retaining the imparted color

Lysochrome Dyes

Oil soluble dyes

Without auxochrome component

To demonstrate fats (histochemical stains)

Sudan Balck B, Sudan IV, Sudan III

Exampled of Lysochrome Dyes

Sudan Black B

Most sensitive Lysochrome dye

Sudan IV

A.k.a Scharlach R

For neutral lipids, staining them red

Sudan III

First Sudan dye introduced to histochemistry

Good fat stain for CNS tissue