From: vanburen%flovax.dnet@rocdec.roc.wayne.edu
Date: Wed Nov 03 1993 - 12:48:08 EST
David L. Abbott asked about sorters. We were using our FACS Vantage to sort CD8+ lymphocytes from mouse spleen. Our starting percentage ranged from 20% to 40%. One of the goals was to collect as many cells as possible (for further study). Using beads, I determined that the best flow rate for this experiment was 7000/second (tested 1000/sec to 10000/sec). This rate offerred the maximum of cells collected per time. On a bad day, purity would drop to almost 98%, but most sorts produced 99% pure populations. Obviously, your starting conditions are different, so you might want to investigate the best strategy for your conditions. I believe most people (and I may be wrong here) would use a two-pass strategy: an enrichment followed by a "clean-up" (purity sort). The FACS Vantage has built-in features (sorting modes) for both of these methods: ENRICH and NORMAL. A magnetic-bead sorter (or column or other method) may also be used for enrichment. You may want to contact Glenn Paradis at MIT. He has a lot of sorting experience, and knows other operators that have experience sorting fetal cells. Glenn may be reached at (617)253-6454. /\/\/\_ Eric Van Buren, vanburen%flovax.dnet@rocdec.roc.wayne.edu \ \ \ Immunology & Microbiology \_^_/ Wayne State University
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