Re: Viability of FACS sorted cells

From: Simon Monard (smonard@trudeauinstitute.org)
Date: Fri Mar 23 2001 - 16:46:51 EST


Hi

What sheath fluid are you using, some commercial sheath fluid is rather toxic to cells,
I use the Biosure preservative free stuff and am pleased with it. You can just make up
some PBS and use that. Are you giving the cells some protein before and after sorting,
BSA before and FCS after, FCS can make cells clumpy so is often avoided before sorting.

Good luck



Simon Monard
FACS Lab Manager
Trudeau Institute
Saranac Lake
NY12983

Ph 518 891 3080 X352


>>> Gilbert Pitts <grpitt2@pop.uky.edu> - 3/23/2001 9:10 AM >>>

Hi Everyone,

I have recently tried to sort GFP-identified cells from the mouse
suprachiasmatic nucleus.  Unfortunately,  the cells appear to be dead
after the FACS sort is completed.  I've been using a trypsin dispersion
is HBSS (supplemented with Hepes and pen/strep).  Does anyone have any
ideas of how I can increase cell viability?  Thanks in advance!

Gilbert

--
Gilbert Pitts, Ph.D.
Post-doctoral Scholar
Dept. of Physiology
University of Kentucky
800 Rose St, Room MS508
Lexington, KY 40536-0084
Tel. (859) 257-1645
Fax: (859) 323-1070



This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:01:12 EST