The preparation of cultured cells of vascular origin for transmission electron microscopy

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Abstract
A method is described for obtaining optimal, reproducible ultrastructure of vascular smooth muscle cells and vascular endothelial cells in culture. Routinely grown cultures are prepared for TEM with a precise regimen of fixation, postfixation, en bloc staining, dehydration, and embedment. The most important aspects of this procedure are the following: (1) fixation with a percentage‐gradient series of glutaraldehyde solutions at 37°C, (2) immediate postfixation with osmium tetroxide solution, and (3) block‐staining with uranyl acetate solution to eliminate any extraction of constituents during subsequent processing.