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Nigrosin is a mixture of synthetic black dyes (CI 50415, Solvent black 5) made by heating a mixture of nitrobenzene, aniline and aniline hydrochloride in the presence of a copper or iron catalyst. Its main industrial uses are as a colorant for lacquers and varnishes and in marker-pen inks. Sulfonation of nigrosin yields a water soluble anionic dye, nigrosin WS (CI 50420, Acid black 2).
In biology, nigrosin is used for negative Staining of bacteria. 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 like methylene blue or carbol fuchsin, 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: living cells exclude the dye, but it enters dead cells.
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