Hi Folks, A colleague here at UCONN Health describes the following problem, I am trying to perform a simple sort of cells positive for GFP expressing 4T-1 cells - a breast cancer cell line. My problem is the cells agglutinate to each other severely in suspension. As a result the sorting has a unstable system threshold - rapidly fluctuating between 2K and 5K with a consequent very high abort rate (50-80%). The buffer I used to resuspend the cells is serum-free DMEM with 2% BSA and 1mM EDTA. The cells rapidly agglutinate in this buffer, so rapidly that you can visualize it by holding the tube to the light. Even in trypsin solution the cells tend to aggregate to each other if not being vigorously dispersed. I would appreciate any suggestions on how to resuspend this tumor cell line and keep them suspended. Thanks, Tao Hu Sibtech, Inc. Newington, Ct.
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