Selective Staining of Hepatitis B Surface Antigen in Thick Epoxy Sections of Liver

Hepatitis B surface antigen (HB2Ag) in epoxy-embedded liver tissues can be stained by aldehyde-fuchsin stain. Sections are oxidized in KMnO4 acidified with H2SO4, then bleached in NaHSO3, both at 70 C. Heating for oxidation and bleaching are absolutely necessary. Diluted aldehyde-fuchsin stain adjusted to pH 1.5 to 1.8 with NaOH is used for staining. HBsAg is specifically stained purple. Other components such as mitochondria and bile pigments are also stained, but are easily distinguished from HBsAg. This staining method is advantageous for the identification of HBsAg-positive cells for electron microscopic observation.