The tungstate-stabilized tetramethylbenzidine reaction for light and electron microscopic immunocytochemistry and for revealing biocytin-filled neurons.

摘要: A peroxidase reaction product that can be easily distinguished from standard diaminobenzidine (DAB) products is needed for pre-embedding electron microscopic double-antibody labelling studies. Benzidine dihydrochloride (BDHC) and gold-substituted silver reactions are unsatisfactory double because they lack sensitivity reliability and/or compromise ultrastructure. We show here light immunocytochemistry done with a modification of the tungstate-stabilized tetramethylbenzidine (TMB) (Weinberg Van Eyck 1991) which yields crystalline product. With this method, we have obtained excellent immunolabelling variety antigens, including tyrosine hydroxylase, enkephalin, serotonin, Fos protein retrogradely transported cholera toxin B subunit (CTB). The TMB-tungstate useful ultrastructural crystals contrast well amorphous reactions. more sensitive reliable than benzidine gives better ultrastructure reaction. also neurons filled biocytin by intracellular injection visualized either (LM) or (EM) microscopy.