© 1995 Oxford University Press
RESEARCH-ARTICLE |
Ultra-sensitive FISH using peroxidase-mediated deposition of biotin- or fluorochrome tyramides
Department of Cytochemistry and Cytometry, Medical Faculty, Leiden University Wassenaarseweg 72, 2333 AL Leiden, The Netherlands
*To whom correspondence should be addressed
Received November 7, 1994; Revised January 13, 1995; Accepted January 13, 1995
We describe a detection principle for indirect fluorescence in situ hybridization (FISH) methods that with only one or two antibody layers dramatically improves FISH signal intensities. The method uses as a first layer an anti-hapten immunoglobulin [or (strept)avidin] conjugated to peroxidase. The quintessence of the method is the use of fluorochrome- or biotin labelled tyramides as peroxidase substrates to generate and deposit many fluorochrome or biotin molecules close to the in situ bound peroxidase may either be directly evaluated under the fluorescence microscope or after another incubation with fluorochrome-labelled (strept)avidin.
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