Electron microscopic immunoperoxidase staining of insulin using 4-chloro-1-naphthol after osmium fixation.
Ultrastructural localization of insulin in B cells of guinea pig pancreas was accomplished after osmium fixation with an immunoperoxidase procedure that utilized 4-chloro-1-naphthol (CN) in the substrate solution. The principal features of this protocol were: a) osmium tetroxide postfixation; b) omission of hydrogen peroxide "etching"; c) use of CN instead of diaminobenzidine in the substrate solution; d) elimination of osmium tetroxide after the substrate reaction; e) uranyl acetate and lead citrate counterstaining. This procedure produces intense specific staining with low background using highly dilute antiserum, and appears to be useful for postembedding immunoperoxidase staining of a variety of peptide antigens in osmium-fixed tissue.
D000818:Animals; D006168:Guinea Pigs; D007124:Immunoenzyme Techniques; D007328:Insulin / Q000032:analysis*; D007515:Islets of Langerhans / Q000648:ultrastructure*; D008297:Male; D008854:Microscopy, Electron; D009284:Naphthols; D009993:Osmium Tetroxide; D013194:Staining and Labeling
D007328:Insulin; D009284:Naphthols; C025917:4-chloro-1-naphthol; D009993:Osmium Tetroxide