[Cytometry] monocyte-platelet aggregates

Fent János stjano at freemail.hu
Thu Apr 25 09:10:50 EDT 2013


Dear Adeeb,

The rate of the spontaneous complex formation  between platelets and leukocytes depends on the type of the anticoagulant applied. According to our experience citrate as anticoagulant is the best choice since in heparinized blood the complex formation is very rapid, while there is no complex formation in EDTA.

Furthermore, When your whole blood sample contains optimal number of leukocytes for the flow cytometric measurement, the platelet concentration is so high that the coincidence of platelets and monocytes can not be neglected, resulting in an overestimation of their complexes. How to to overcome this effect experimentally, please, see our publications on this topic:

Bihari P et al: An easy-to-use practical method to measure coincidence in the flow cytometer--the case of platelet-granulocyte complex determination. J Biochem Biophys Methods. 2008 Apr 24;70(6):1080-5.

Fent J et al: Impact of coincidence on granulocyte-platelet complex determination by flow cytometry is evaluated by a novel computer simulation model of coincidence. J Biochem Biophys Methods. 2008 Apr 24;70(6):1086-90.

We wish you good luck to your projects,
Susan Lakatos
and János Fent


>Adeeb Rahman <adeeb_r at yahoo.com> írta:
>
>    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
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