Hallo Barbara, I've been sorting SL2 cells for many years now, started off on the Vantage, nowadays on a MoFlo. I have never come across viability problems. I use 40psi, 70um nozzle and FacsFlow sheath fluid. If you are sure about the medium, then I would suggest changing the nozzle as it may be damaged and somehow shearing your cells. regards Ann At 15:57 09.11.01 -0500, you wrote: >We have made several attempts to sort a Drosophila cell line, SL2 >(Schneider cell line 2). We have been experimenting with >non-transfected wild-type cells. > >Based on the FSC vs SSC dot plot, we start out with 95% of the cells >in the viable cell region. After sorting, very few cells are whole; >most have been turned into debris. We are examining the sorted cells >under a microscope and re-running them within 5 minutes of their >being sorted. The problem is not the sort medium as unsorted cells >remain viable when diluted in either medium or sheath fluid. > >I have turned down the pressure (FACS Vantage SE with TurboSort) to 9 >PSI (70 micron nozzle) and have turned down the laser power (argon >only) as low as I can. > >I know that insect cells are very fragile; has anyone sorted these >and maintained viability? > >Thanks >-- >Barbara J Taylor >Facility Manager, FACS Core Lab >Bldg 37 Room 6008 >CCR, NCI, NIH >Bethesda, MD 20892-4255 >phone 301.594.6892 >fax 301.496.8709 >taylorba@pop.nci.nih.gov
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