Differential staining of acid glycosaminoglycans (mucopolysaccharides) by Alcian blue in salt solutions

Summary

The application of the “critical electrolyte concentration” (CEC) concept to the differentiation of acidic glycosaminglycans (mucopolysaccharides) is described. Alcian Blue 8GX stains with increasing selectivity as increasing amounts of magnesium chloride are incorporated into the dye solution. Model experiments with pure polyanions, or artifically carboxylated, phosphorylated and sulphated liver sections, showed that binding of dye to carboxylate or phosphate groups ceased at low electrolyte concentrations (< 0.3M) whereas dye continued to be held by sulphate ester groups at concentrations five to ten times as high. The similarity to the well established cetylpyridinium system for polyanion fractionation is discussed.

Sections of tissues chosen to contain predominantly or characteristically carboxylated mucins, and/or sulphate ester polyanions showed a staining pattern entirely similar to the model sections. Goblet cell mucin in rat ileum stained at < 0.4M MgCl₂, Cartilage at < 0.6M MgCl₂, mast cells at < 0.75M, and corneal stroma at < 1.0M. These results are in agreement with the known contents of sialo-mucin, chondroitin sulphate, heparin and keratansulphate, respectively. The conditions in which this principle can be used in a practical technique are described.