Dear Inter-Flow-ers, I need your help in determining what is going on in a particular experiment. A research colleague is trying to measure sca-1 cells in mice. We look at bone marrow and peripheral blood samples and we use a rat PE IgG2a isotype control to assess the background fluorescence. In the bone marrow samples we see both lymphocytes and granulocytes. When we gate on the lymphocytes and just look at the fluorescence on those cells we get a background of around 6% (leaving the granulocytes in we get a raised background). When we come to look at the peripheral blood, which appears to be mainly lymphocytes, we get a massive background of 66%. Why? I might put it down to none specific binding but why the difference between marrow and peripheral lymphocytes? Any suggestions? Regards, Jeff Barry Flow Operator Paterson Institute Manchester UK
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