Hydrogen peroxide as an oxidizing agent for glycogen demonstration using Schiff’s reagent in rabbit liver fixed with alcoholic and aqueous fixatives

Abstract

ABSTRACT This study investigated the optimal concentration of hydrogen peroxide (H2O2) for demonstrating glycogen in rabbit liver using Schiff’s reaction as compared to the conventional periodic acid-Schiff (PAS) method. Four fixatives, Bouin’s, neutral buffered formalin (NBF), 80% ethanol, and Rossman’s, were used at room temperature (RT) and 4°C. Paraffin sections of rabbit liver (n = 192) were stained using 0.5%, 1% and 2% periodic acid-Schiff (PAS) and 0.5%, 1%, and 2% hydrogen peroxide (H2O2)-Schiff methods. It was found that 0.5% H2O2-Schiff provided good staining results, while 1% PAS produced excellent results for glycogen fixed with 80% ethanol at RT. Also, 0.5% H2O2-Schiff produced good staining of glycogen fixed with 10% NBF at RT as compared to 0.5% PAS which provided satisfactory staining results. Fixation with 80% ethanol at RT produced the best staining quality for glycogen using both oxidizing agents at the optimal concentrations of either 1% periodic acid or 0.5% H2O2.