Hi Barry, The DCs could very well be getting activated by LPS present in your system. Macrophages can also be sensitive to the presence of LPS. There are microtiter plate based tests that you can use to measure LPS levels in your system (we use the LAL QC1000 test from Cambrex). We have been monitoring LPS levels on our MoFlo for over 2 years now and have found the in-line sheath filter to be the greatest source of LPS contamination. For prevention, we take care to dump out whatever is remaining in the sheath tank and rinse it with 70% EtOH, as well as flush the lines with 70% EtOH every evening. We also periodically "soak" the inside of the sheath tank with 5 N NaOH to destroy any LPS present there and change the sample line in addition to the in-line sheath filter. If you need any more details you can contact me off- line. Good luck, Marie Follo Am 5 Apr 2006 um 16:40 hat Barry Moran geschrieben: Hi there- We're sorting cultured murine dendritic cells on a MoFlo (70micron nozzle, 60psi) into culture media. We're finding that the cells appear some-what activated (CD40+, CD80+, CD86+) after the sort compared to unsorted controls. Have many others found this, and if so are there any suggestions or fixes- is it likely to be endotoxins etc in the sheath fluid/lines? Many thanks Barry. Barry Moran Flow Cytometry Facility School of Biochemistry and Immunology 0.19, Biotechnology Building Trinity College Dublin +353 1 608 3575 barry.moran@tcd.ie <http://www.tcd.ie/biochemistry/flow> www.tcd.ie/biochemistry/flow ****************************************************************** Marie Follo, Ph.D. Core Facility Zentrale Klinische Forschung Breisacherstr. 66 Universitaetsklinik Freiburg 79106 Freiburg Germany Tel.: +49 761 270 6377 e-mail: marie.follo@uniklinik-freiburg.de http://www.ukl.uni-freiburg.de/core-facilityReceived on Fri Apr 7 15:58:00 2006
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