Re: H202, Phagocytosis

From: Arnold Pizzey (a.pizzey@ucl.ac.uk)
Date: Mon Dec 20 1999 - 04:28:19 EST


At 15:16 16/12/99 -0500, Joan Kalnitskyyou wrote:
>
>This is not a Flow question, but I thought this group might be able to help
>out.  Does anyone know of any protocols to measure H202 and phagocytosis in
>a manner other than flow cytometry.  We have a PI who has measured both of
>these via flow and is now working in Chile where they have no flow and
>would like to be able to measure these.
>	Any and all input will be greatly appreciated.
>	Thanks in advance,
>	
>	
>
>	Flow Cytometry Lab Supervisor
>	VMRCVM
>	(540) 231-4115
>	FAX 540-231-7367
>	jkalnits@vt.edu


Hello Joan.

Many years ago I tried labelling Candida with fluorescein diacetate (just
grow them up overnight in fda)

On feeding them to neutrophils the neuts acquired a healthy green glow this
was done using the flow. 

 I did also try growing up yeasts in dichlorodihydoflurescein (molprobes)
and giving them to neuts in a fluorimeter cuvette- they generated a
beautiful rising green fluorescence signal. Now, there are some follow-up
experiments that I should have done; e.g. does the observed signal
represent reactive oxygen species generation concomitant to phagocytosis.
As this was just a quick look, I left it there however.

I hope this may be of some use

Best wishes to you (and all) for the holiday season.


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:	+44 171-209-6234
	Fax:	+44 171-209-6222
	email:	a.pizzey@ucl.ac.uk
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