The Influence of Buffers During Fixation on the Appearance of Smooth Endoplasmic Reticulum and Glycogen in Hepatocytes of Normal and Glycogen-depleted Rats

Liver tissue of normal and glycogen depleted rats was prepared for transmission electron microscopy by perfusion fixation and subsequent osmication in the presence of various buffers, dehydration in aethanol and embedding in epon. The use of Na/K-phosphate or Na-cacodylate to buffer glutaraldehyde led to similar appearance and distribution of SER. When Na-cacodylate was used during osmication, more SER membranes were retained but less accumulations of glycogen were found than after osmication in the presence of Na/K-phosphate. Fixation with s-collidine buffered osmium led to an easily recognisable network of SER comprising wide tubules whereas glycogen was hindered to be stained. Veronal acetate or Na-cacodylate supplemented with sucrose resulted in marked dilation and disintegration of SER. A similar effect was obtained when Na/K-phosphate or Na-cacodylate was used in hyposmolar concentration as buffer for glutaraldehyde. Liver of fasted rats or glucagon-treated rats after perfusion with Na/K-phosphate buffered glutaraldehyde and osmication in the presence of Na/K-phosphate or Na-cacodylate comprised glycogen-depleted hepatocytes which contained abundant SER membranes occupying the entire space between other organelles even in samples harvested 3 h after glucagon administration. The diversity in appearance and distribution of SER and glycogen granules, which depends to a large extend on the buffer used, suggests that SER membranes may not be sufficiently stabilized during aldehyde fixation and osmication. We thus consider it likely that large accumulations of glycogen granules are the consequence of disintegration of SER membranes during processing rather than they represent the morphologic substrate of physiological degradation of SER membranes in the course of glycogen synthesis and deposition.