Dear people, we got quite a problem with the measurement of our t-cell clones from mice. The autofluorescence of the unstained cells (no isotype control) is between 100 and 3000 measured in almost all fluorescence channels if we use the calibration of the beads supplied by Becton Dickinson, designed for calibrating for human blood. We calibrated manually (amplification and compensation) and now are not satisfied with "normal" measurement since it is very low now. What I want to know is if you experienced the same problem with mouse-clones (not fresh LNC and also not with murine blood), and if yes, how do you handle it? What range are your amplifiers and compensation, how far do you try to lower autofluorescence measurement, and do you have to calibrate for every new clone? Thanks for the time Stefanie Reuss MZ Nervenheilkunde - Neurologische Klinik mit Poliklinik Rudolf-Bultmann-Str.8, 35039 Marburg, Germany, Fax +49 6421 28 7055, Phone +49 6421 28 5480
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