Re: CFSE analysis by flow

From: Aki Hoji (akhst7@imap.pitt.edu)
Date: Thu Aug 29 2002 - 08:19:53 EST


Jose,

It is possbile to use 4-color comb on XL.  I used to do IFNg-PE, CD3-ECD,
and 69-PC5.  Other combs are also possible.  Obviously, this all depends on
how strong cells are labeled with CFSE.  If you have money to invest, you
should get a PC7 filter kit, and this will make you life so much easier.

If you still want to voltage setting, 4-color comp, and CFSE-labeling for 4
color, please contact me.


Aki Hoji
University of Pittsburgh
Dept. Infectious Diseases and Microbiology
Rm 526 Parran Hall, GSPH-IDM,
130 Desoto Street,
Pittsburgh, PA 15261
Tel: 412-624-3072
Fax: 412-624-4953


  
> 
> From: Jose Benito <jbenito1@hotmail.com>
> Date: Mon, 26 Aug 2002 14:49:51 +0200
> To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu>
> 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
> 
> 
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