Open AccessCombined beta-galactosidase and immunogold/silver staining for immunohistochemistry and DNA in situ hybridization.
Author(s) -
W van den Brink,
Chris van der Loos,
Herman Volkers,
R Lauwen,
Femke van den Berg,
H. J. Houthoff,
Pranab K. Das
Publication year - 1990
Publication title -
journal of histochemistry and cytochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.971
H-Index - 124
eISSN - 1551-5044
pISSN - 0022-1554
DOI - 10.1177/38.3.1689335
Subject(s) - immunogold labelling , staining , microbiology and biotechnology , immunohistochemistry , in situ hybridization , silver stain , biology , in situ , epitope , monoclonal antibody , dna , chemistry , antibody , biochemistry , gene , gene expression , genetics , immunology , organic chemistry
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.
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