Ultrastructural Localization of Cholesterol in Human Arteriosclerosis
Several reports have appeared recently demonstrating a technique for the localization of cholesterol in a variety of tissues. The technique is based on the premise that free cholesterol reacts with digitonin to form cholesterol digitonide which appears as needle-like crystals or spicules under the electron microscope.In order to study the fine structure of the aorta, surgical biopsies from several patients (ranging in age from 45 to 68 years) and with clinical evidence of arteriosclerosis were fixed promptly in Flickinger's fixative (2% formaldehyde, 2.5% glutaraldehyde) containing 0.2% digitonin but buffered with .1M sodium cacodylate. After fixation for 3 hours aortic samples were washed in cacodylate buffer and post fixed in buffered osmium tetroxide for 2 hours & embedded in Epon-Araldite.