Scanning electron microscopy in the evaluation of endothelial integrity of the fatty lesion in atherosclerosis.

The luminal surface of fatty lesions of atherosclerosis was viewed by scanning electron microscopy (SEM). Endothelial cells were outlined by staining intercellular junctions with silver and the aortas were fixed in situ at physiological pressure. When aortas were dehydrated by passage through organic solvents followed by critical point drying from liquid CO2, there was considerable disruption of the luminal surface and it was not possible to correctly interpret the morphological integrity of the endothelium. In contrast, simple air-drying of aortas, without solvent dehydration after fixation, allowed the integrity of the cell layer overlying the lesion to be evaluated. The success of this technique was attributed to the retention of arterial lipids during dehydration of the tissue.