This study investigated the optimal concentration of hydrogen peroxide (HO) 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 (HO)-Schiff methods. It was found that 0.5% HO-Schiff provided good staining results, while 1% PAS produced excellent results for glycogen fixed with 80% ethanol at RT. Also, 0.5% HO-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% HO.
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