Hi Andy, I have done something very similar quite recently. The cells had been tagged with a GFP variant that would not survive the ethanol fixation required to enable me to use PI so we went back to the reliable Hoechst technique. After disaggregation the cells were stained with 10µg/ml Hoechst 33342 for 20 minutes at 37¡C. As the staining was performed elsewhere, the cells remained in the dye until they were analysed but this didnt affect the viability as judged by PI exclusion. The Hoechst fluorescence was collected between 390 and 480nm and the GFP using a 510/20bp filter. Good luck! Derek On Thu, 26 Aug 1999, Andrew Morschauser wrote: > We have a researcher that would like to look at GFP and cell cycle in > drosophila disks using PI or Hoechst. Does anyone out there have a > labeling protocol for cells containing GFP for cell cycle? > Andy Morschauser > University of Pennsylvania Cancer Center > Flow Cytometry & Cell Sorter Shared Facility ************************************************************************ Derek Davies Voice: (44) 0171 269 3394 FACS Laboratory, FAX: (44) 0171 269 3100 Imperial Cancer Research Fund, e_mail: derek.davies@icrf.icnet.uk London, UK Web Page: http://www.icnet.uk/axp/facs/davies/index.html In tenebris lux *************************************************************************
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