Immunogold technique applied to simultaneous identification of T6 and HLA-DR antigens on Langerhans cells by electron microscopy

Abstract

A double-labeling immunogold technique in electron microscopy and specific monoclonal antibodies to surface antigens of Langerhans cells (OKT6 and BL2) were applied to assess directly the coexpression of two cell surface antigens (T6 and HLA-DR antigens) in a heterogeneous epidermal cell suspension. Electron microscopic examination of double-labeled cells revealed that all Birbeck granule-containing Langerhans cells bound OKT6 and BL2. The preparation of markers with colloidal gold particles and the procedure for double labeling are described. Several problems related to the steric hindrance and current artifacts are illustrated by micrographs and also discussed.