Dual staining methods for carbohydrates using lectin-gold silver techniques.

Three dual staining methods were established for the histochemical detection of saccharide residues and acidic groupings of carbohydrates in light microscopy. The first method consisted of combined lectin-gold silver (LT-G-S) and alcian blue (AB) pH 1.0 techniques, whereas the second staining technique was composed of LT-G-S and AB pH 2.5 procedures. These two techniques were found to color saccharide residues and acidic groupings of carbohydrates in black and blue shades respectively, which exhibited a high contrast between the both. The third methods, LT1-G-S-LT2-PO-DAB techniques, yielded reaction products of blackish and yellowish brown shades, which represented the localizations of two different saccharide residues in one and the same section. According to the results of the experimental and control procedures, the present three dual staining methods are believed to be reliable, reproducible and unusually useful for light microscopic histochemical studies on acidic and non-acidic carbohydrates.