Hi Tina If I were you I would run one of your samples on a benchtop instrument to see if anything strange happens. If they look "normal" on a scan or calibur I would put them back on the Vantage and try bocking the second and third lasers to see if its a curious laser cross-talk problem. Have you looked at the freshly isolated cells on the Vantage? If they are the same as your loaded cells then it points to a misaligned laser. Fresh lymphocytes are very small compared to cultured cells so do require much higher PMT voltages but you should be able to get the negatives nicely on screen. Your cells could be clumping which may account for the strange side scatter. Simon Monard Trudeau Institute Algonquin Avenue Saranac Lake NY12983 518 891 3080 X162
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