Re: Clumping cells

From: Tom Mc Closkey (thomasm@nshs.edu)
Date: Tue Oct 24 2000 - 11:14:09 EST


Jeff Barry wrote:
>
>> A colleague is trying to look for apoptosis using the sub G1, Propidium
> Iodide, method however, his cells stubbornly refuses to form single cells
> and continue to aggregate causing our FacsCalibur to block every time we put
> the samples on. Can anyone relieve our misery by suggesting any sample
> preparation tips that will prevent cell clumping.
>

	A variation on this assay utilizes hypotonic lysis to isolate
individual nuclei instead of analyzing whole cells.  It is described in:
J Immunol Meth, 139: 271-279, 1991.  This should eliminate cell/cell
binding.

Good luck,
Tom

*****************************************************************************
Thomas W. Mc Closkey, Ph. D.
Director of Flow Cytometry, North Shore University Hospital
Assistant Professor of Pediatrics, New York University School of
Medicine
Boas Marks Biomedical Research Center, 350 Community Drive
Manhasset, Long Island, New York 11030
ph:  516-562-4844 [office], 516-562-1135/4641 [lab]  fax:  516-562-2866
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