staining dyes Staining is a technique used to enhance contrast in samples, generally at the microscopic level. Stains and dyes are frequently used in histology (microscopic study of biological tissues), in cytology (microscopic study of cells), and in the m ...

, nigrosin (CI 50415, Solvent black 5) is a mixture of black

synthetic dyes A dye is a colored substance that chemically bonds to the substrate to which it is being applied. This distinguishes dyes from pigments which do not chemically bind to the material they color. Dye is generally applied in an aqueous solution and ...

made by heating a mixture of nitrobenzene,

aniline Aniline is an organic compound with the formula C6 H5 NH2. Consisting of a phenyl group attached to an amino group, aniline is the simplest aromatic amine. It is an industrially significant commodity chemical, as well as a versatile starti ...

, and

hydrochloric acid Hydrochloric acid, also known as muriatic acid, is an aqueous solution of hydrogen chloride. It is a colorless solution with a distinctive pungent smell. It is classified as a strong acid Acid strength is the tendency of an acid, symbol ...

in the presence of

copper Copper is a chemical element with the symbol Cu (from la, cuprum) and atomic number 29. It is a soft, malleable, and ductile metal with very high thermal and electrical conductivity. A freshly exposed surface of pure copper has a pinkis ...

iron Iron () is a chemical element with Symbol (chemistry), symbol Fe (from la, Wikt:ferrum, ferrum) and atomic number 26. It is a metal that belongs to the first transition series and group 8 element, group 8 of the periodic table. It is, Abundanc ...

. Related to

induline Induline is a dye of blue, bluish-red or black shades. Induline consists of a mixture of several intensely colored species, so the name is often indulines. It was one of the first synthetic dyes, discovered in 1863 by J. Dale and Heinrich Caro. Th ...

, it is a mixture of

phenazine Phenazine is an organic compound with the formula (C6H4)2N2. It is a dibenzo annulated pyrazine, and the parent substance of many dyestuffs, such as the toluylene red, indulines, and safranines (and the closely related eurhodines). Phenazine ...

-based compounds. Its main industrial uses are as a

colorant A colourant/colour additive (British spelling) or colorant/color additive (American spelling) is a substance that is added or applied in order to change the colour of a material or surface. Colourants can be used for many purposes including printing ...

for

lacquers Lacquer is a type of hard and usually shiny coating or finish applied to materials such as wood or metal. It is most often made from resin extracted from trees and waxes and has been in use since antiquity. Asian lacquerware, which may be ca ...

and

varnishes Varnish is a clear transparent hard protective coating or film. It is not a stain. It usually has a yellowish shade from the manufacturing process and materials used, but it may also be pigmented as desired, and is sold commercially in various ...

and in

marker pen A marker pen, fine liner, marking pen, felt-tip pen, felt pen, flow marker, sign pen (in South Korea), vivid (in New Zealand), texta (in Australia), sketch pen (in South Asia) or koki (in South Africa), is a pen which has its own ink source ...

inks.

Sulfonation Aromatic sulfonation is an organic reaction in which a hydrogen atom on an arene is replaced by a sulfonic acid functional group in an electrophilic aromatic substitution. Aryl sulfonic acids are used as detergents, dye, and drugs. Stoichiometry a ...

of nigrosin yields a water-soluble

anionic dye Acid dyes are anionic, soluble in water and are essentially applied from acidic bath. These dyes possess acidic groups, such as SO3H and COOH and are applied on wool, silk and nylon when ionic bond is established between protonated –NH2 group of ...

, nigrosin WS (CI 50420, Acid black 2).

Uses

Aside from its main use in lacquers, small amounts of nigrosin are used for

negative staining In microscopy, negative staining is an established method, often used in diagnostic microscopy, for contrasting a thin specimen with an optically opaque fluid. In this technique, the background is stained, leaving the actual specimen untouched ...

bacteria Bacteria (; singular: bacterium) are ubiquitous, mostly free-living organisms often consisting of one Cell (biology), biological cell. They constitute a large domain (biology), domain of prokaryotic microorganisms. Typically a few micrometr ...

. as well as the capsule-containing fungus ''

Cryptococcus neoformans ''Cryptococcus neoformans'' is an encapsulated yeast belonging to the class Tremellomycetes and an obligate aerobe that can live in both plants and animals. Its teleomorph is a filamentous fungus, formerly referred to ''Filobasidiella neoforman ...

''. The shapes and sizes of the organisms are seen as color-free outlines against the dark background. An advantage of using this method, rather than regular

positive stains Staining is a technique used to enhance contrast in samples, generally at the microscopic level. Stains and dyes are frequently used in histology (microscopic study of biological tissues), in cytology (microscopic study of cells), and in the ...

like methylene blue or

carbol fuchsin Carbol fuchsin, carbol-fuchsin, carbolfuchsin, or Castellani's paint (CAS ) is a mixture of phenol and basic fuchsin that is used in bacterial staining procedures. It is commonly used in the staining of mycobacteria because it has an affinity fo ...

, is that prior fixation by heat or alcohol is not needed, so the organisms are seen in more lifelike shapes. Furthermore, negative staining with nigrosin can reveal some microorganisms that cannot be stained by regular methods. Nigrosin WS is used in tests for viability because living cells exclude the dye, but it enters dead cells.

References

{{Reflist Azin dyes Staining dyes