danielg@uncmed.med.unc.edu (11/09/90)
We need to isolate *viable* human monocytes from whole blood, with >90%
purity, and highest possible yield, of course :-). In the past we have
used mono-poly resolving media (Flow Labs), which gives two bands, one
containing polynuclear cells, the other containing mononukes. We need to
remove the lymphocytes (we have also had a slight problem with some pmn
contamination in the mono layer - poly layer, however, has been quite
pure).
All suggestions warmly welcomed. Email or post.
Thanks in advance,
dan
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S A V E T H E W H A L E S , K I L L O U R U N B O R N ??
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Daniel G. Sinclair - UNC Chapel Hill - Satisfied in Jesus. :-). Ahhhhhh.kelley@aclcb.purdue.edu (Steve Kelley) (11/10/90)
In article <1573@beguine.UUCP>, danielg@uncmed.med.unc.edu writes: > >We need to isolate *viable* human monocytes from whole blood, with >90% >purity, and highest possible yield, of course :-). In the past we have Have you tried flow cytometry? Purity > 90% should be no problem at all, but what exactly do you mean by highest possible yield? Flow will limit you to looking at a couple thousand cells per second, and picking out whatever fraction of the input are monocytes. If flow cytometry alone won't get you the numbers you need, perhaps a two-step procedure will do it? I believe Larry Arnold is at UNC, in the dept of Micro and Immuno. He can fill you in on all the gory details of sorting. Steve Kelley Name: Steve Kelley Internet: kelley@aclcb.purdue.edu Phone: 317 494-8638