Radioiodinated cholesteryl iopanoate, a nonhydrolyzable cholesteryl ester probe, showed increased uptake into atherosclerotic aortas of cholesterol-fed rabbits in comparison with normal rabbits. Auto-radiography of the aortas showed the radioactivity to be concentrated in areas of visible atherosclerotic involvement. Lipid extraction and thin-layer chromatography of this tissue as well as liver, adrenal, and plasma confirmed the resistance of this probe to hydrolysis. These findings suggest that 125I-cholesteryl iopanoate may prove useful for noninvasively monitoring atherosclerosis in intact laboratory animals.