Jose,
CFSE being so bright on autoflourescence will make life very
difficult for you to realize practical compensation values for your
protocols. Try and optimize the PMT bias for CFSE alone and bring it as low
as possible on FL1 Voltage gain. You will be happy with the other colors and
most likely you'll get manageable compensations values before you exhaust
your cell volumes to acquire.
Thanks.
Peter Mucheru.
-----Original Message-----
From: Jose Benito [mailto:jbenito1@hotmail.com]
Sent: Monday, August 26, 2002 3:50 PM
To: cyto-inbox
Subject: CFSE analysis by flow
Hi flowers, I am trying to work out a protocol for acquisition of cells
labelled with CFSE and 3 more colors (PE, ECD and PE-Cy5). These are PBLs
stimulated with mitogens and antigens. The problem id that CFSE gives a
staining so bright (in the fourth log decade) that everything else
(specially PE) is completely contaminated by the strong CFSE signal. I tried
last friday to compensate for PE channel without succes (I run out of cells
to acquire). I am going to try again today. Does anyone have any experience
working with CFSE and other colors? Is tehre anyway to find out a
cytosettings for this combination?
Many thanks
Jose Miguel Benito
Madrid
Spain
_____
Join the world's largest e-mail service with MSN Hotmail. Click Here
<http://g.msn.com/1HM1ENXX/c157??PI=31901>
This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:26:20 EST