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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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