Hi Flowers, I'm a novice in cell sorting and trying to sort eosinophils for further culture. The problem is the viability of the sorted eos which is clearly lower than that of unsorted cells. We shurely have to replace the FACSFlow we use as sheathfluid by PBS in the future. But we also tried to reduce the sample pressure. Our sorter is a Cytomation MoFlo which usually runs at 60.2 psi sample pressure and 60.0 psi sheath pressure. When reducing the pressure, we were told, the difference between sample and sheath pressure should be larger than on high pressures to prevent the sample tube from filling with sheath fluid and to get beads or cells to the nozzle at all. The lowest sample pressure which we didn't get a reflow with was about 58 psi when we lowered the sheath pressure down to1 psi. In the mailing I list I found that many people are sorting at about 30 psi, but I don't know if this is sample or sheath pressure. So, my questions are: 1. What pressure is critical for after-sorting viability: sample or sheath? 2. If it is sample pressure, how do I get to 30 psi? 3. Has anyone sorted eos and further cultivated these cells? Is pressure critical at all? Thanks in advance Martin Förster Inner Medicine IV / Pneumology Friedrich-Schiller-University Erlanger Allee 101 07740 Jena / Germany Tel.: +49 (0)3641 939 580 FAX: +49 (0)3641 939 572 e-mail: martin.foerster@med.uni-jena.de
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