Dear Andrew, I have done GFP & cell cycle work, but not with drosophila disks. I suggest using Hoerchst for the following reasons. 1/ If your GFP is not bound to organelles or structures within the cell it will be lost when you puncture the cell wall to let the PI in. I have seen this using a Laser Scanning Cytometer, I was able to read the GFP signal, then stain the same cells with PI & overlay the 2 results. Virtually all the GFP was lost in the few minutes it took to perform the PI staining. 2/ GFP can be sensitive to pH changes within the cell, ie its fluorescence can be reduced or quenched by cell death or some fixation techniques. I have not seen this in relation to cell cycle specifically, but have seen it in sorted samples. Hope this helps. Regards Rob W. At 14:03 08/26/1999 +0100, you wrote: >Hi All, >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? >Thanks, >Andy Morschauser >University of Pennsylvania Cancer Center >Flow Cytometry & Cell Sorter Shared Facility R. Wadley, B.App.Sc. M.L.S, Grad.Dip.Sc.MM Laboratory Manager Cellular Analysis Facility School of Microbiology & Immunology UNSW, New South Wales, Australia, 2052 Ph (BH) +61 (2) 9385 3517 Ph (AH) +61 (2) 9555 1239 Fax +61 (2) 9385 1591 E-mail r.wadley@unsw.edu.au www http://www.micro.unsw.edu.au/caf.html
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