Dyes are molecules used to stain the structures of animal and plant tissues, that is, they provide colors to cell structures and extracellular matrix. There are a broad variety of dyes used in histological techniques, and they are selected depending on the sample and the features to be study. Dyes are selected regarding the color they provide, molecular weight, solubility, their affinity to bind and react with tissular molecules.
The structures to be stained and the color the dyes may provide can be guessed from their chemical nature. Regarding the electrical charge, which can be deduced from the molecular composition, dyes are classified into cationic or basic dyes, which stain nuclei and acid carbohydrates, and anionic or basic dyes that stain the cytoplasm and the extracellular matrix. The non-charged dyes can be molecules that stain a variety of structures, liposoluble dyes for staining lipid depots, and mordants that color myelin and nuclei. The size of the dye molecules and their ability to form aggregates is sometimes important because of their difficulty to diffuse through tissues. For example, anionic dyes with different sizes can be selectively used to stain different structures.
Diversity of dyes (modified from Kiernann 2009).
Name | Molecule | Type | Color | Staining | Procedure |
Iodine | Inorganic | Blue | Chromosomes and starch and glycogen depots. Gram bacteria. Protozoa. | Lugol's solution | |
Osmium tetroxide | Inorganic | Dark grey, bblack | It is a histochemical reaction: lipids, proteins, Golgi apparatus, nerves, neurons, glia. | Golgi method, Marchi method. | |
Gold and silver salts | Inorganic | Impregnations and toning. | |||
Prussian blue | Inorganic | Blue | Cystein and cystin | Histochemical method for cystein. | |
Naphthol green | Nitroso | Complex metal -dye |
Green | Collagen | Mixed with anionic dyes. |
Picric acid | Nitro | Anionic | Yellow | Cytoplasm | Thricromic stainings, it is a mordant. |
Martius yellow | Nitro | Anionic | Yellow-orange | Cytoplasm | Together with anionic gyes. |
Orange G | Azo | Anionic | Orange | Cytoplasm | Papanicolaou, Mallory trichrome, Heidenhain trichrome. |
Metanil yellow | Azo | Anionic | Yellow | Cytoplasm, connective tissue | Anionic contrast. |
Biebrich scarlet | Azo | Anionic | Scarlet | Collagen, basic proteins | Masson trichrome (acid fucshin replacement) |
Amaranth | Azo | Anionic | Red | Cytoplasm, nuclei | Gabe thrichrome |
Bisckmarck Brown | Azo | Cationic | Brown | celular extensions | |
Janus green B | Azo / azine | Cationic | Green | Mitochondria, bacteria | |
Alcian yellow | Azo /thiazole | Cationic | Yellow / fluorescent | Bacteria | |
Procion brilliant red M2B | Azo | Reactive | Red | Vital staining for teeth and bone. | |
Sudan IV | Azo | Solvent | Red | Lipid droplets | |
Oil red O | Azo | Solvent | Red | Lipid droplets | |
Black Sudan B | Azo | Solvent / Cationin |
Black | Lipd droplets | |
Congo red | Azo | Direct | Red | Amyloid depots / pH indicator | |
Benzo blue BB | Azo | Direct | Blue | Collagen fibers (birefringense) | |
Chlorazol black E | Azo | Direct | Black | For plants, fungi and microorganisms. | |
Sirius red F3B | Azo | Direct | Red | Collagen fibers (birefringence) | |
Auramine O | Arylmethane | Diarylmethane | Yellow | Tuberculosis bacillus | |
Pararosanilina | Arylmethane | Aminotriarylmethane | Schiff reagent | ||
Basic fuchsin | Arylmethane | Aminotriarylmethane | Red | Nucelus | Schiff reagent |
Rosaniline | Arylmethane | Aminotriarylmethane | Magenta | Non commercial | |
Magenta II | Arylmethane | Aminotriarylmethane | Magenta | NoN coMmercial | |
Violet crystal | Arylmethane | Aminotriarylmethane | Violet | Gram + bacteria | |
Methyl green | Arylmethane | Aminotriarylmethane | green-blue | ||
Ethyl green | Arylmethane | Aminotriarylmethane | Green-blue | Nucleus, DNA, RNA | |
Acid fuchsin | Arylmethane | Aminotriarylmethane, anionic | Red | ||
Fast green FCF | Arylmethane | Aminotriarylmethane, anionic | Green-blue | Connective | |
Aniline blue (methylen blue) | Arylmethane | Aminotriarylmethane, anionic | Blue | Connective | |
Coomassie brilliant blue | Arylmethane | Aminotriarylmethane, anionic | Blue | Proteins in gels | |
Sodium fluorescein | Xanthene | Anionic | Fluorescent | Proteins | |
Eosin | Xanthene | Anionic | Pink | Connetive, cytoplasm | |
Pyronine Y | Xanthene | Cationic | Red | DNA | |
Rhodamine B | Xanthene | Cationic | Fluorescent | ||
Oregon green | Xanthene | Cationic | Fluorescent | ||
Acriflavine | Acridine | Cationic | Yellow, Fluorescent | DNA | |
Acridine orange | Acridine | Fluorescent | DNA, carbohydrates, vital | ||
Neutral red | Azin | Cationic | Red | Nuclei, lysosomes, vacuoles, vital dye | |
Safranin O | Azin | Cationic | Red | plant lignin, nucleus | |
Azocarmine G y B | Azin | Anionic | Red | Cytoplasm | |
Cresyl violet | Oxazin | Cationic | Violet | Nucleus | |
Gallocyanin | Oxazin | Cationic | Green-blue | DNA | |
Nile blue | Oxazin | Anionic | Blue | Vital dye | |
Orcein | Oxazin | Cationic | Red | Elastin, chromosomes | |
Thionin | Thiazin | Cationic | Blue | ||
Azure A, B y C | Thiazin | Cationic | Blue | Blood | |
Methylen blue | Thiazin | Cationic | Blue | ||
Methylen blue | Thiazin | Cationic | Green-blue | Nucleus | |
Tolouidin blue | Thiazin | Cationic | Blue, metachromatic | ||
Methylene Violet (Bernthsen | Thiazin | Cationic | Blue | Blood smears | |
Calcofluor white | Polyene | Fluorescent | Cellulose | ||
Indigo | Carbonyl | Anthraquinone | |||
Alzian red | Carbonyl | Anthraquinone | |||
Nuclear fast red | Carbonyl | Anthraquinone | |||
Pale Turquoise Sirius blue | Phthalocyanine | ||||
Alcian blue 86 | Phthalocyanine | ||||
Cuprolinic blue | Phthalocyanine | ||||
Luxol fast blue | Phthalocyanine |
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Bibliography ↷
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Kiernann JA. (2009). Histologican and histochemical methods. Theory and practice. 4th edition. Scion Published Ltd. ISBN 9789048422
Llewellyn B. (retrieved: 2022). Stain files. http://stainsfile.info/dyes/dyes.htm
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