Re: Annexin V (AV) and propidium iodide (PI) staining to identify apoptotic/necrotic cells

From: Martin Kelly (M.G.Kelly@queens-belfast.ac.uk)
Date: Thu Feb 10 2000 - 13:22:01 EST


Thanks for your reply. The numbers are not usually too high 
(probably less than 10% total gated events) but is higher in 
some individuals, maybe moreso in normals, strangely. Someone 
else suggested cell nuclei (described as 'bare nuclei') also, as 
a possibility. My only contention with this is that one might 
not expect to find nuclei in the area on the FS v SS plot on 
which we have gated. We tend to gate in an area of high FS and 
SS, and i believe that nuclei wouldn't lie there. What do you 
think?

		Martin Kelly

On Thu, 10 Feb 2000 10:13:57 -0800 "Dennis J. Young" 
<djyoung@ucsd.edu> wrote:

> I vote for cell nuclei without PS-containing membranes. Never bothered to
> sort them to verify this. Are there high numbers in your samples?
> 
> 
> At 12:29 PM 2/9/00 +0000, you wrote:
> >
> >Dear friends,
> >we would like to ask some advice r.e. the interpretation of some 
> >work we have been doing on induced sputum and BAL from normal 
> >subjects and COPD patients. We have been using 'flow' to look at 
> >the granulocytes, and examine the AV and PI staining in the 
> >events which we have gated on, which we feel are in the region 
> >in which one would expect to find granulocytes. Cells coming up 
> >AV+PI- we interpret as apoptotic; AV+PI+ as necrotic. This would 
> >seem to be conventionally how it is desribed in papers we have 
> >looked at. However, we were not sure how one might interpret 
> >cells which are AV-PI+? Are these just cells in an advanced 
> >state of necrosis? Is there another interpretation?
> >We value any advice.
> >With best wishes. yours sincerely,
> >
> >					Martin Kelly
> >
> >P.S. since drafting this email, I have had a look through the 
> >excellent email archives, and found some commentary on this 
> >issue, but I don't think that the issue was ever really 
> >resolved. Any definitive opinions or references would therefore 
> >be appreciated.
> >
> >----------------------
> >Martin Kelly
> >Department of Clinical Biochemistry, 
> >Queen's University of Belfast,
> >Institute of Clinical Science,
> >Grosvenor Road,
> >BELFAST. BT12 6BJ
> >Tel No: +44 (0)28 9026 3267
> >Fax No: +44 (0)28 9023 6543
> >Email: m.g.kelly@qub.ac.uk
> >
> >
> >
> >
> >
> 
> Dennis J. Young
> Mail:<<mailto:djyoung@ucsd.edu>>
> WWW:<<http://cancer.ucsd.edu/SResources/flow.htm>>
> New Location & New Area Code, too!
> (since June, 1999)			Voice : (858) 822-0407
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> University of California, San Diego
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> 9500 Gilman Drive La Jolla  CA  92093-0671

----------------------
Martin Kelly
Department of Clinical Biochemistry, 
Institute of Clinical Science,
Grosvenor Road,
BELFAST. BT12 6BJ
Tel No: (01232) 263267



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