Cheryl, Concerning fluorescent in situ hybridization for the detection of mRNA species by flow cytometry, sensitivity is a major problem. In situ RT-PCR has not worked well. We have used our enzymatic amplification staining (EAS) technology for this problem and have achieved excellent results (attached 2 powerpoint figures). Flow-Amp's EAS kit for intracellular antigens with FITC conjugated primary antibodies was used with substitution of a FITC conjugated cDNA probe for Epstein-Barr Virus EBER1 RNA instead of an antibody. The difficulty in the procedure was getting the hybridization conditions right but the results were impressive. David David Kaplan, MD, PhD Professor of Pathology Case Western Reserve University Co-Founder, Flow-Amp Systems Hello All, I've had a query from one of our facility users regarding the ability of flow cytometry to detect single FISH probes. The most recent posting from the group that I can find is from 1996 and I'm wondering if everyone has simply decided that flow is not sensitive enough for this. It was mentioned that incorporation of digoxigenin into the PCR reaction apparently increased single to noise ratio, but is it really enough to enable detection? Any input would be greatly appreciated. Thanks, Cheryl
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