[Histonet] (no subject)

From:"Garry Ashton"

Message: 2

Date: Tue, 30 Nov 2004 13:21:51 -0500

From: Susan Q Wells 

Subject: [Histonet] Frozen cell pellet

To: Histonet@lists.utsouthwestern.edu

Message-ID: <41ACBA3F.9030602@bms.com>

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Does anyone have advise on how to freeze cells that have been spun down
in a micro centrifuge tube? In the past I've snap frozen the tube,
loosened the cells by flicking the tube and then embedded the pellet in
OCT. The cells look OK, but the cells in the middle don't look as good
as the ones on the perimeter.Any advise would be appreciated.


Sue Wells


Hi Sue,

When I freeze cell pellets down that have been rinsed in PBS I remove
them from the ependorf using a wide pasteur and place them in OCT, in
effect to remove the PBS.

I place them in a second OCT "bath" for a futher rinse and then snap
freeze in more OCT on a chuck.

I think the important thing is firstly they are not to densley packed,
and secondly the PBS is replaced by OCT.

Hope this helps.





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