From: Yoav Altman (yoav@pobox.com)
Date: Tue Sep 16 2003 - 21:15:11 EST
Flowers, An investigator bringing me samples for calcium flux studies has some questions about her data that I'm not able to answer. I'd appreciate any help from the calcium wizards out there. We're running Jurkat cells loaded with Indo-1 on a FACSVantageSE DiVa. I'm using 80mW of MLUV from a Krypton laser for excitation and BD's 510SP to split the signal to my two UV detectors. UV1 and UV2 have 405/20 and 424/44 filters respectively; nothing unusual, as this all came from BD's calcium filter set. Voltages for the PMT's are 580 (UV1) and 520 (UV2). When we stimulate the cells with OKT3 we see a nice increase in violet (Calcium bound Indo-1) emission. The problem is that we don't see a corresponding decrease in blue (unbound) fluorescence. Do any of you have any ideas as to why we aren't seeing a decrease in unbound Indo-1 fluorescence? Thanks, Yoav Dot plots of cells before and after stimulation with OKT3. Time-course of the same data set. -- Yoav Altman Manager, High Throughput Cell Analysis Core The BURNHAM INSTITUTE 10901 North Torrey Pines Road La Jolla, CA 92037 Voice: (858) 646-3100 xFLOW (x3569) Fax: (858) 646-3167 email: <yoav@burnham.org>
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