RE: P19 cell death

From: Fischer, Randy (NIAMS) (fischer1@mail.nih.gov)
Date: Sun Mar 24 2002 - 20:39:34 EST


Gene,

We are not and have not had this problem with our SE.  Perhaps it is the
CFSE treatment or the cell lines.  Have you run the cell line and sorted for
just size or for a different marker?

Randy T. Fischer
NIH/NIAMS
Autoimmunity Branch
Building 10, Rm 6D57
9000 Rockville Pike
Bethesda, MD 20892
(301) 594-3537
fischer1@mail.nih.gov

> ----------
> From:		Pizzo,Eugene
> Sent:		Thursday, March 21, 2002 4:48 PM
> To:	Cytometry Mailing List
> Subject:	P19 cell death
>
>
> Folks,
>
> I'm seeing a fantastic amount of cell death following a low speed/low
> pressure sort on my VantageSe and am wondering if anyone has experience
> with these cells in particular or tips for preventing cell death in
> general.
> I'm getting about 10-20% live cell recovery following a typical low
> pressure sort at 3K/sec,  8 psi, and 18khz with a 100um nozzle.
> I normally get an average 70% recovery across the board.The cells are P19
> embryonic carcinoma
> cells with CFSE as a marker.	They are collected in DMEM with 20% FBS
> and pen/strep.
> Thanks.
>
> Gene Pizzo/UCONN Health
>
>



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