Automated detection of RNA FISH by imaging flow cytometry.

<p>(A) Organization of the MMTV-Ras transgene in the 3134 cell line. The 3134 cell line contains approximately 200 tandem repeats of MMTV-LTR driving viral Ras RNA on chromosome 4. (B) A Dig-labeled DNA probe was generated by nick translation of the pM18 plasmid containing the viral Ras coding sequence. Agarose gel electrophoresis revealed a DNA probe size within a 250–500 base pair range. (C) Imaging of cells following probe hybridization showed the presence of distinct nuclear FISH signals at a 40X magnification. The FISH signal was detected with anti-Dig secondary antibodies conjugated to a green-fluorescent dye. BF = Bright Field; DRAQ5 = nuclear stain.