Re: Embyonic Stem Cells

From: Ann Atzberger (Ann.Atzberger@EMBL-Heidelberg.de)
Date: Thu Dec 07 2000 - 04:35:10 EST


Hi Chris,

unless there's a good reason for it, I would suggest washing just once; the
more you manhandle cells the more you compromise the viability. Instead of
resuspending the cells in PBS/BSA you could try medium ( will however give
you increased autofl. which will not be a problem if you have a good signal
above that for the stained population).Also sort into medium.
The settings for the Vantage are fine,(I've used a 70um nozzle with no
adverse effects) unless of course you have a bad nozzle.

regards
Ann




At 09:49 05.12.00 -0800, you wrote:
>
>We are having problems with the viability of our Embryonic Stem Cells
>after sorting.  I am using a VantageSE at 9psi, 21K ddf, and a 100u
>nozzle.  Prior to sorting, these cells have been stained, spun, and
>washed 5 times.  Is anyone successfully sorting this type of cell??
>
>chris
>
>trussell@gnf.org
>
>



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