Polyacrylamide films as a tool for investigating qualitative and quantitative aspects of the staining of glycosaminoglycans with basic dyes

摘要: With the introduction of model films polyacrylamide gel into which purified glycosaminoglycans (GAGs) have been ‘incorporated’, direct recording metachromatic spectra with virtually no interference corresponding orthochromatic peaks has become possible. Because this system yields situations comparable to those stained sections under microscope, it is well suited for investigating qualitative and quantitative aspects histochemical staining procedures. Previous experiments shown that aqueous conditions only minor differences can be observed between different GAGs complexed a suitable dye (e.g. Toluidine Blue O, Thionin, Safranin Cresyl Violet, Crystal Violet). In non-aqueous media, such as glycerol ethylene glycol, complexes O revealed special pattern heparin, having peak (517 nm) about 30 nm lower than all other GAGs. This observation formed basis method microspectro-photometric detection heparinin situ was worked out by combining film microspectrophotometric data obtained from rat mast cells. Since limited number cells necessary obtaining reliable method, presence heparin in cytoplasmic granules normal human basophilic granulocytes could thus proved directly.