Variations in kidney ultrastructure with a variety of paraformaldehyde fixation techniques

Variations in fine structure of glomeruli and proximal tubules were studied to determine the results of several fixation techniques with paraformaldehyde. Immersion fixation techniques generally resulted in poor fixation, while the quality of fixation by perfusion was directly related to the concentration of the paraformaldehyde. The best technique was fixation by perfusion and immersion in 3% paraformaldehyde for 2 h followed by a post-fixation in 1% OsO4 for 1 h. Intracellular edema was noted in all epithelial, mesangial and proximal tubular cells that were from poorly fixed kidneys. Swollen mitochondria and the dilated endoplasmic reticulum were common in many of those cells. Mesangial and proximal tubular cells were the first to swell. Poor fixation produced the early stages of autolysis. The criteria for good paraformaldehyde fixation are discussed.