Re: absolute cell counts using FACS

From: Arnold Richard Pizzey (a.pizzey@UCL.ac.uk)
Date: Tue Jun 19 2001 - 02:39:39 EST


>Hi everybody ,
>Can anyone recommend a microsphere kit for performing simple cell counts
>on numerous samples using the FACS? BD's Truecount beads are only sold
>along with a MAb. Our cells do not have to be labelled. Any simpe
>alternative? (NB: our good ol' Coulter particle counter is dead)
>Tx
>Karim

Hello Karim,

I have to do absolute cell counts here sometimes: I take a bottle of flow
check beads (Beckman-Coulter) and wash them in FCS X2 then resuspend them
at 1exp6/ml in FCS. Then you just spike your sample with them, the only
thing that you have to take some care over is the initial count; I count
~1000 beads on a haemocytometer. If you don't need them sterile you can add
~1.0% azide.

Regards

Arnold

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	Arnold Richard Pizzey
	Department of Haematology
	Royal Free and University College London Medical School
	98 Chenies Mews
	London WC1E 6HX
	U.K

	voice:	+44 020-7679-6234
	Fax:	+44 020-7679-6222
	email:	a.pizzey@ucl.ac.uk
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