Comparison and analysis of the staining methods for cholesterol

Cholesterol staining is a useful approach for the visualization, localization and quantification of cholesterol in cells or tissues, which is frequently used to investigate the mechanisms of some diseases such as arteriosclerosis, Niemann-Pick disease type C, and Alzheimer's disease. It can be accomplished through various microscopes including light microscope, fluorescent microscope, and electronic microscope. During the past decades, various types of methods for cholesterol staining with different principles have been established for different applications. It is important to choose an appropriate method that is suitable for particular experimental aims, features and conditions. At present, three kinds of methods are frequently applied: filipin fluorescent method, BCθ(a biotinylated and carlsberg protease-nicked derivative of perfringolysin O) toxin method, and cholesterol oxidase-diaminobenzidine(oxidase-DAB) method. Four kinds of methods are scarcely applied: Schultze method, perchloric acid-naphthoquinone method(PAN), digitonin method, and o-phthalaldehyde method. In this review, the principles, advantages, and disadvantages of these methods are compared with the emphasis of the application, sensitivity, and specificity.