Tissue cholesterol preservation: Factors associated with retention of cholesterol in rat sciatic nerve fixed for electron microscopy

摘要

AbstractThe retention of cholesterol in rat sciatic nerve fixed in glutaraldehyde‐digitonin was investigated. Maximal retention (>85% ) was achieved after fixation at room temperature, post‐fixation in OsO4, and dehydration in an ethanol series, 60, 80, 95, 100% (2 changes), five minutes per change. Fixing in the cold, increasing the dehydration time, changing from ethanol to acetone, or reducing the concentration of digitonin from 0.2% to 0.02%, resulted in a decrease in cholesterol retained. The greatest loss of cholesterol was found in the two changes of absolute ethanol, and therefore we recommend dehydration through 95% ethanol, followed by infiltration and embedding according to the procedure of Idelman ('