An enzymatic method for the histochemical localization of free and esterified cholesterol separately

An enzymatic method for the histochemical localization of cholesterol is presented. It makes possible the localization of free cholesterol, cholesterol esters, or both and is compatible with routine histological staining procedures. The method is based on the production of H2O2 from free cholesterol by cholesterol oxidase. Sites of peroxide production are visualized by a brown reaction product formed in a peroxidase-catalysed reaction between diaminobenzidine and H2O2. Cholesterol esters can be demonstrated as cholesterol after hydrolysis by cholesterol ester hydrolase. Some examples of the application of the method are given.