Demonstration of Neutral Polysaccharides with Fluorescence Microscopy using Acridine Orange

Abstract

ACRIDINE orange, a basic fluorochrome dye, stains the basophilic constituents of cells, such as nucleic acids (De Bruyn et al.¹, Morthland et al.², Armstrong³) and acid mucopolysaccharides (Hicks and Matthaei⁴, and Kuyper⁵), but aqueous solutions of this dye do not stain glycogen, paramylum and polysaccharides of the cyst wall of protozoa. All these polysaccharides can be stained with acridine orange after sulphation. Several sulphation techniques have been tried, and the best results have been obtained with chlorosulphonic acid in dry pyridine (Kramer and Windrum⁶). For the sulphation of glycogen 5 min treatment at 65° C is enough, whereas paramylum bodies and cyst wall require 10–15 min treatment at 70°–75° C.