Biological stains Active coloring subst: acid component

a. Natural dyes Inactive base: sodium salt of a sulfonate

b. Synthetic or artificial dyes of rosaniline
Picric acid: fix, differentiates, stains;
Natural dyes counterstain; decalcifier, softener
a. Hematoxylin TCA, PA, chromium fixed tissues
b. Cochineal dyes 2. Basic dyes
c. Orcein Basic component that combines with acid
d. Saffron radical: H2SO4, acetic or HCl
Methylene blue
Hematoxylin Fixatives: Mercuric chloride,
Hematoxylin campechianum formaldehyde
Most valuable, powerful n+c, polychromatic Acidic cell structures: basophilic
Natural ripening: oxidation of hematoxylin 3. Neutral dyes
Artificial ripening: chemical oxidation using Acid+basic dyes
strong oxidizing agents: H2O2, K2MnO4, Na Ethanol or acetic acid fixed tissues
perborate, Na iodate
Over ripening: excessive oxidation Common stains
1. Hematoxylin
Cochineal dyes Most common
Coccus cact Mordants: Alum and Iron
Treated with alum carmine Lakes or colored complexes
Treated with picric acid picrocarmine Aluminum salt lakes: blue
neuropathological studies Ferric salt lakes: blue black
Treated with aluminum chloride Bests a. Aluminum hematoxylin
carmine stain glycogen Progressive or regressive
Blueing
Orcein b. Ehrlichs hematoxylin
Regressive
Vegetable dye from lichens ammonia
Acid decalcified tissue
blue or violet colors
Not ideal for frozen tissues
Elastic fibers
15 to 40 minutes
Litmus indicator
c. Harris hematoxylin
4% glacial HAc: nuclear stg
Synthetic dyes or Coal Tar dyes Regressive
Hydrocarbon benzene or C6H6 Stable for 6 months
Aniline dyes Routine, exfoliative cyto, sex
Chromophores: definite atomic groupings chromosomes
visible colors 5 to 20 minutes
Chromogens: simple benzene compounds d. Coles hematoxylin
no permanent color In sequence with Celestine blue
Auxochrome: imparts permanent color to 10 minutes
chromogens by electrolytic dissociation e. Mayers hematoxylin
Regressive or progressive
Nuclear cytoplasmic glycogen
1. Acid dyes MPS
 Chloral hydrate preservative Basic Fuchsin
3 to 6 months AFS, mitochondria, smooth muscles
f. Iron hematoxylin Feulgens and Schiffs: adlehyds
Differential or regressive Van Giesons: connective tissues, mucin,
Acid Alcohol: differentiator elastic
Mordants: a. Carbol Fuchsin
Weigerts solution: ferric ammonium b. Colemans Feulgen
chloride; muscle fibers, connective tiss c. Schiffs
Heidenhains solution: ferric ammonium d. Mallorys
sulfate (iron alum; regressive, n+c: e. Aldehyde/Gomoris
chromatin, chromosomes, nucleoli,
centrosomes, mitochondria OTHER STAINS
g. Phosphotungstic acid
Progressive Bismarck Brown Contrast: Grams
Paraffin, celloidin, frozen AFS, Papanicolau,
12 to 24 hours diphtheria
Carmine Chromatin
Eosin Aluminum Cl: Best Carmine
Differential Mayers Basic dye: Acidic stain
Routine counterstain after hematoxylin Carmalum
and before methylene blue Celestine Blue Fixed sxns
Background stain Congo Red Axis cylinders in embryos
a. Yellowish or Eosin Y Crystal Violet Amyloid in frozen sxns and
Most common; 15 seconds to 3 mins platelets in blood
b. Bluish Giemsa stain Leukocytes
Deeper red Gold sublimate Metallic impregnation
c. Ethyl eosin Gold and mercuric chloride
Eosin S, eosin alcohol soluble Iodine Oldest; starch granules
Rarely used: Eosin S, Eosin B Lugols iodine Glycogen, amyloid, corpora
amylacea
Acid Fuchsin-Picric Acid (Van Giesons stain) Janus Green B Mitochondria
Picric acid + acid fuchsin Malachite green Contrast: ascaris eggs,
Connective tissues erythrocytes, bacterial
a. Acridine orange spore, decolorizer,
Dead vs living cells counterstain
Green fluorescence for DNA and red Methyl green Chromatin green
fluorescence for RNA Methylene blue Polychroming, plasma
cells, fresh sputum
Methylene Leukocytes: reddish purple
violet
b. Acridine red 3B
Neutral red Granules, vacuoles of
Deposits of calcium salts, phosphatase
phagocytic cell
activities
Night blue Carbol fuchsin substitute:
c. Alcian blue
AFS
Acid MPS
Orcein Elastic: Taenzer Unna
Connective tissue, mucin Orcein, dermatologic, skin
 fibers 10ml aniline water + to 1 L of hot
Osmium Fat: osmium tetroxide to distilled H2O
tetroxide osmium dioxide black 4. Phenol
Picric acid Contrast to acid fuchsin: Aqueous: 0.5-5%
connective tissue
Counterstain to CV
Tissue fixative, decalcifier
Prussian blue Paints, circulatory system
by injection
Rhodamine B Osmic acid: blood and
glandular tissues
Silver nitrate Spirochetes, reticulum,
fibers
Toluidine blue Nissl granules,
chromophilic bodies
Victoria blue Neuroglia
Lipid stains
Sudan Black Most sensitive
Phospholipids, neutral fats
Sudan IV TG: neutral lipids
Sudan III CNS

Chief solvents
1. Water
distilled
2. Alcohol
Ethanol
Absolute methanol: blood
3. Aniline water