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Combined beta-galactosidase and immunogold/silver staining for immunohistochemistry and DNA in situ hybridization

W van den Brink, C van der Loos, H Volkers, R Lauwen, F van den Berg, HJ Houthoff and PK Das

Department of Pathology, Immunohistochemical, Academic Medical Center, Amsterdam, The Netherlands.

A combination of beta-galactosidase enzyme and the immunogold/silver staining method was studied for evaluation of double-staining experiments. Applications are shown for immunohistochemical double staining using two monoclonal antibodies and for combined immunohistochemistry and DNA in situ hybridization in one tissue section. The following advantages for the present double-staining method were evaluated: superior sensitivity of the immunogold/silver staining method for at least one epitope, which also allows detection of biotinylated DNA probes. The structure of the indolyl precipitate after revelation of beta-galactosidase activity did not show a concealing effect during a sequential double-staining method, as compared with the visualization of peroxidase with diaminobenzidine. These factors, and the sharply contrasting colored reaction products of beta-galactosidase (blue-green) and the immunogold/silver staining method including silver enhancement (brown-black), allow clear distinction of mixed-stained cell constituents.

Volume 38, Issue 3, pp. 325-329, 03/01/1990
Copyright © 1990 by The Histochemical Society


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