Histochemical procedures for the simultaneous visualization of neutral sugars and either sialic acid and its side chain O-acyl variants or O-sulphate ester. I. Methods based upon the selective periodate oxidation of sialic acids.

Abstract

Five new methods, based upon the selective oxidation of sialic acid residues with 0.4 mM periodic acid in approximately 1 M hydrochloric acid at 4 degree C for 1 h (PA), have been devised for the simultaneous visualization of neutral sugars and either sialic acid and its side chain O-acyl variants or O-sulphate ester. In the first of these, the selective periodate oxidation-borohydride reduction-saponification-selective periodate oxidation-Thionin Schiff-saponification-borohydride reduction-periodic acid-Schiff (PA-Bh-KOH-PA-T-KOH-Bh-PAS) technique, sialic acids with O-acyl substituents at C7, C8 or C9 (or which have two of three side chain O-acyl substituents) stain blue while neutral sugars with periodate-sensitive vicinal diols (hexose, 6-deoxyhexose, and N-acetylhexosamine) stain magenta. The second method, the saponification-selective periodate oxidation-Thionin Schiff-saponification-borohydride reduction-periodic acid-Schiff (KOH-PA-T-KOH-Bh-PAS), stains all sialic acids blue and neutral sugars magenta. In the third procedure, the selective periodate oxidation-Thionin Schiff-borohydride reduction-periodic acid-Schiff-saponification (PA-T-Bh-PAS-KOH) method, sialic acids without side chain substituents (or which have an O-acyl substituent at C7) stain blue and neutral sugars stain magenta. In the fourth method, the saponification-selective periodate oxidation-borohydride reduction-Alcian Blue pH 1.0-periodic acid-Schiff (KOH-PA-Bh-AB1.0-PAS) technique, O-sulphate esters stain aquamarine blue and neutral sugars stain magenta. In all of these techniques mixtures of the components stain in various shades of purple.(ABSTRACT TRUNCATED AT 250 WORDS)