[Cytometry] monocyte-platelet aggregates

Gary Warnes g.warnes at qmul.ac.uk
Fri Apr 19 10:29:13 EDT 2013


Dear Adeeb,
	When looking at platelets you should use sodium citrate as an
anti-coagulant (discarding the first ml of activated blood) and also it
would be better if you did not lyse the RBCs as this will also affect
platelet numbers. To get around the high RBCs count load the sample with
50nM LDS751 or DRAQ5 to gate and threshold on nucleated material - Prof
Macey at The Royal London Hospital has quite a few papers on looking at
platelet-cell aggregates.

Hope this helps

Gary

Dr Gary Warnes
Flow Cytometry Manager 
Flow Cytometry Core Facility
Blizard Institute 
Barts and The London School of Medicine and Dentistry
London University
4 Newark Street 
London
E1 2AT
Tel No.0207-882-2402
Fax No. 0207-882-2180
Website http://www.icms.qmul.ac.uk/flowcytometry/index.html

-----Original Message-----
From: cytometry-bounces at lists.purdue.edu
[mailto:cytometry-bounces at lists.purdue.edu] On Behalf Of D. Robert
Sutherland
Sent: 19 April 2013 15:02
To: Adeeb Rahman
Cc: Cytometry at lists.purdue.edu
Subject: Re: [Cytometry] monocyte-platelet aggregates

Adeeb,
I am not an expert in tis area, but I believe EDTA is the anti-coagulant of
choice for the types of studies you are doing.
HTH
best
rob

D. Robert Sutherland
Toronto General Hospital/University Health Network

On 2013-04-18, at 4:11 PM, Adeeb Rahman wrote:

> Dear flow folk,
> 
> Do any of you have experience detecting monocyte-platelet aggregates by
flow? I've been trying to quantify MPAs using antibodies against CD14 (clone
HCD14) and CD42b (clone HIP1) to stain whole blood, followed by RBC
lysis/fixation with BD FACSLyse.Using this protocol, I'm finding that a very
high frequency of monocytes are CD42b+ (over 90%), which is much higher than
the frequency reported in various publications (typically ~10%). 
> 
> 
> I've looked at blood collected in citrate tubes and sodium heparin tubes
and have looked at tubes drawn later in the draw sequence (i.e. not the
first tube drawn) but am still seeing the same thing. Does anyone have any
thoughts about what may be causing this high frequency of MPAs? 
> Thanks,
> 
> Adeeb
> 
>  
> ___________________________
> 
> 
> Adeeb Rahman
> Postdoctoral Fellow
> Division of Liver Diseases
> Mount Sinai School of Medicine
> New York, NY
> _______________________________________________
> Cytometry mailing list
> Cytometry at lists.purdue.edu
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