Hi Mark et al, We had similar experience sorting mouse CNS cells in a StarPlus. We concluded that the cells simply don't survive all the kicking around, and especially don't like sitting in a pool of saline after being sorted. Our best results were sorting into about 3mL of medium, and changing the collection tubes every 5 minutes or so to minimise the time the cells spend in the saline layer on top of the medium. This stuff was done by Andrew Ford & Jon Sedgwick, can probably find references if you need. Good Luck, Joseph. At 04:39 5/11/99, Mark A. KuKuruga wrote: >Hi, all . . . >I'm sorting sciatic nerve cells, specifically neural stem cells as a >target. We find that the sort recovery (actual count vs. instrument >count) is often less than 70%, perhaps as low as 50%. The optimal >breakoff has been verified using 10 micron particles to be accurate, and >we see much higher recoveries with other cell types. >This has been done on a Vantage SE, either high-speed or at low >pressures, using sort envelopes of 1, 1.5, 2, and 3 drops. Threshold >rate is ~300-600 cells/sec, so we're not talking high aborts. I can >only speculate that these cells are not traveling uniformly in the >stream . . . >Anyway, any ideas? Joseph Webster, Flow Cytometry Facility, Centenary Institute Locked Bag No.6, Newtown, NSW 2042, AUSTRALIA. Ph: 61-2-9565-6110 Fax: 61-2-9565-6101 e-mail: J.Webster@centenary.usyd.edu.au
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