[Histonet] Single-cell susension from tissue

[anh2006 <@t> med.cornell.edu]

What enzymes are you using for digestion? Just curious .... Have you looked into the BD Medimachines for cell dissociation? They work great .... So easy!

-----Original Message-----
From: Mark Tarango <marktarango <@t> gmail.com>

Date: Tue, 08 Jul 2008 20:43:24 
To: <hyters <@t> onid.orst.edu>
Cc: <histonet <@t> lists.utsouthwestern.edu>
Subject: Re: [Histonet] Single-cell susension from tissue


Hi Stephen

You can vortex but I'd keep them in the enzyme bath longer.  If you're
worried about losing a population of cells (or the epitopes getting chewed
up from digestion), remove the cells that have come out of the tissue and
hold them in PBS.  You can put the remaining tissue back for digestion (you
don't want to exclude a population of cells because they never came out of
the tissue).  Different populations of cells can come out at different times
and  you can even end up selecting out populations based on how fast you are
centrifuging.

Mark

On Tue, Jul 8, 2008 at 7:50 PM, <hyters <@t> onid.orst.edu> wrote:

> hello, having trouble fully dissociating dermal tissue to use in a cell
> count on a hemacytometer.  Can I vortex cells in suspension or should I just
> keep them in the enzyme bath a bit longer? (worried about surface markers
> for FACS)
>
> thanks for any help,
> stephen
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet