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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