The effect of fixatives and temperature on the quality of glycogen demonstration

Abstract

Glycogen is demonstrated in a number of lesions and is diagnosrically significant, particularly in certain tumors. To stain glycogen accurately, it is essential to choose a suitable fixative, temperature and staining method. We used rabbit liver to assess these variables. Specimens were fixed in three fixatives at two temperatures: 10% formalin, neutral buffered formalin (NBF) and Bouin's solution at 37 and 4°C. Seventy-two paraffin sections were prepared and stained with periodic acid-Schiff (PAS), hexamine (methenamine) silver and Best's carmine methods. Negative control sections using diastase digestion were used for all methods to confirm the presence of glycogen. For the PAS reaction, Bouin's fixative gave better results at both temperatures compared to the other fixatives. For hexamine (methenamine) silver, the quality of staining was improved for tissues fixed in both 10% formalin and NBF at 37°C compared to Bouin's solution. Both 10% formalin and NBF at 4°C gave better results than Bouin's solution. For Best's carmine, Bouin's solution gave the best results for tissues fixed at 4°C. Fixation of tissues with NBF at 37°C gave the best quality staining. We concluded that the quality of glycogen staining in paraffin sections is greatly affected by both the fixative and the temperature of fixation.