Combination of non-isotopic in situ hybridisation with detection of enzyme activity, bromodeoxyuridine incorporation and immunohistochemical markers.

The advent of non-isotopic in situ hybridisation allows the possibility to detect the presence of both mRNAs and other markers in cells. We have established conditions for simultaneous analysis of gene expression and a variety of other immunohistochemical markers in tissue sections. We report the analysis of expression of a family of transcription factors (Sox genes) in combination with detection of: (1) protein antigens (using both monoclonal and polyclonal antibodies); (2) bromodeoxyuridine to mark cells which are proliferating; and (3) acetylcholinesterase activity. The approaches we describe, which demonstrate the compatibility of non-isotopic in situ hybridisation with a range of other treatments, should be generally applicable and open to many variations of probe, antibodies and colour detection systems.