抄録
Qualitative and quantitative techniques for the histochemical localization of cholesterol in the aorta were developed. It is based on the same principle as conventional enzymatic determination of total cholesterol in serum by the so-called diformazan stain method. The intimal fiat region of the 5th intercostal aorta were obtained from human adults upon autopsy. The aorta were fixed in 10% formalin and were frozen at -120° under liquid N2 gas delivered through a special nozzle. The aortic slices were prepared using a glass knife and a JB-4 automicrotome. The sections were laid out on the glass slide and were exposed to overhead drops of 1% Triton ×-100 solution and were then stained using an enzymatic total cholesterol reagent (Nikon Shoji). The localization of the cholesterol was observed by microscope, after which 70 points of the percent of extinction in the aortic slices was measured by scanning using a microspectrometer set at a 535nm. To compare the present histochemical method with biochemical cholesterol determination, the same aortic samples were extracted by CHCl3-MeOH (2:1). The total cholesterol in the aliquots was determined by an enzymatic method. Results by the two methods were similar. The advantages of the histochemical method for total cholesterol include direct deternlinatioll of cholesterol. An assay was also possible with observation of localization and distribution of total cholesterol.
