Reply below Posted for general viewing.... > --------------------- Forwarded Message Follows - - - - - - - >Hi- > >I hope this is the proper route to post something on the archive. I've >tracked down several protocols for propidium staining, but I do have one >question they do not address. Don't you need to centrifuge the stained >samples before running them through the flow cytometer? I would >imagine that you still have some PI in solution, and that it would give >you an awfully high background signal. Any comments? > >Thanks. This is a wonderful forum. > >Bob Crow >crow@cbnmr.tamu.edu >---------------------------------- Bob, We routinely use PI staining for live/dead discrimination. It does not need to be washed out prior to running. In fact, PI will leach out if washed. We add it to a tube, wait a few seconds, and then run the sample on our FACSCAN. If you are doing PI for DNA where the cells are permeabilized, I would imagine the same routine applies as PI is reversible in its binding to DNA. I am sending a copy of this to Paul in hopes he will have it posted for the group's info. Good Luck, Randy Fischer NIH/NIA ---------------------------------------------------------------------- posted by J.Paul Robinson, Purdue University Cytometry Labs Professor of Immunopharmacology robinson@flowcyt.cyto.purdue.edu PH:765-494 6449 FAX:765-494 0517 web http://www.cyto.purdue.edu
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