Immunohistochemical staining using coupled immunofluorescence and immunocomplex technique

An immunomicroscopic method combining the use of fluorescent antibodies and enzyme immunocomplex technique is presented. The basic steps of this procedure are: 1. Rabbit antiserum (IgG) to the antigen to be determined, 2. FITC-conjugated antirabbit globulin, 3. Peroxidase + antiperoxidase complex, 4. 3,3-diaminobenzidine + H2O2. The method renders it possible to study simultaneously in the same section antigens and antibodies that are to be localized, and that both by immunofluorescent and immunoenzymatic techniques, it provides means for further elimination of some of the deficiencies of immunofluorescent technique, and in suitable cases it also makes electronoptic examination available.