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Studies in fluorescence histochemistry: I. The demonstration of sulphomucins *
Author(s) -
STOWARD PETER J.
Publication year - 1967
Publication title -
journal of the royal microscopical society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0368-3974
DOI - 10.1111/j.1365-2818.1967.tb04507.x
Subject(s) - lithium aluminium hydride , chemistry , thionyl chloride , fluorescence , saponification , staining , hydrolysis , ferric , alum , feulgen stain , hydride , nuclear chemistry , chromatography , inorganic chemistry , organic chemistry , biochemistry , chloride , medicine , dna , physics , quantum mechanics , pathology , hydrogen
SYNOPSIS Sulphated mucosubstances in sections of coagulant‐fixed tissue emit a variable yellow to red fluorescence after being stained with purified coriphosphine at pH 2 to 4. Unfortunately this fluorescence is sometimes indistinguishable from a similarly coloured fluorescence emitted by nuclei, oxidized sulphoproteins and some sialo‐mucins, which, however, can be eliminated more or less selectively either by (1) a preliminary Feulgen hydrolysis and condensation with N,N‐dimethyl‐m‐phenylene diamine; or (2) by pretreatment with a solution of ferric alum; or (3) by a previous methylation with methanolic thionyl chloride, followed by reduction with lithium aluminium hydride in hot dioxane and then by saponification (a sequence of reactions which, in theory but not always in practice, should remove all polyanionic groups in the tissue except sulphate groups on sulphated mucopolysaccharides); or (4), best of all, by staining sections in a very dilute solution of thiazol yellow at pH 2 after they have been stained in coriphosphine. The rationale and limitations of these methods are discussed critically.