Standardization of grocott`s methenamine (hexamine) silver method for glycogen demonstration in liver tissue

Abstract Purpose Demonstration of glycogen can be done in different lesions and is considered diagnostically significant, mainly in some tumors. Glycogen staining is affected by the type of fixative, the temperature of fixation, and the staining technique. Methods Grocott`s methenamine (Hexamine) silver technique quality was assessed after 4 different types of fixatives at 2 different temperatures (Bouin's solution, 10% neutral buffered formalin (NBF), 80% alcohol, and Rossman's solution at room temperature (RT) and 4°C; for 24 hours). These variables were studied to optimize this technique for glycogen demonstration. Archived paraffin blocks were used in this study. They were prepared from one rabbit`s liver and thirty-two paraffin sections were prepared and stained with Grocott`s methenamine (hexamine) silver technique. Results Eighty percent ethanol gave better results at both RT and 4°C in comparison to the other fixatives. Conclusion 80% alcohol at 4°C seems to provide the best staining results for glycogen with the Grocott methenamine (hexamine) silver technique at the level of this study.