RE: Jurkat and sorting

From: Newsom, Brian S. (BSNEWSOM@txccc.org)
Date: Thu Jun 17 1999 - 14:39:07 EST


Nadia,
We have sorted Jurkats with a 70um tip on a MoFlo with no problems. Most
people say your tip should be 3X the diameter of the cell or greater.
Therefore, the 70um tip can sort cells up to at least 23um and in
practicality, probably to at least 35um without any problems, as long as the
cells are nice ones (i.e. don't tend to clump during sorting and tend to
stay in rough spheres in suspension instead of elongating etc.). The other
variable is the sheath pressure. Cells that easily clog a system at 13 psi
do not tend to clog nozzles so badly at 30 or 60 psi. I would try to sort
them using a 70um tip and if it does not work go to a 100um tip.

Brian Newsom
Director, Flow Cytometry
Center for Cell and Gene Therapy
Baylor College of Medicine

-----Original Message-----
From: NADIA.MASTROIANNI.NM@bayer-ag.de
[mailto:NADIA.MASTROIANNI.NM@bayer-ag.de]
Sent: Wednesday, June 16, 1999 10:27 AM
To: cyto-inbox
Subject: Jurkat and sorting




I have a very naive question:
I'm in the process of starting a new project using Jurkat cells and I would
sort them using a BD FACSVantage.
I've just read an article and in the experimental procedure they write that
the
flow cytometer was equipped with pulse processing and MacroSort flow cell.
Is
it really necessary to sort Jurkat cells? I thought that MacroSort was
useful
for sorting plant cells or pancreatic islets!

Has someone experience in sorting Jurkat cells? Any particular advice?

Nadia




This archive was generated by hypermail 2b29 : Wed Apr 03 2002 - 11:53:37 EST