| Standardization of platelet-derived microparticle counting using calibrated beads and a Cytomics FC500 routine flow cytometer: a first step towards multicenter studies? | |
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MedLine Citation:
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PMID: 18983485 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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BACKGROUND: Platelet microparticles (PMPs) have proved useful to identify patients with vascular risk. However, PMP counting, which is currently done by flow cytometry (FCM), needs to be standardized. OBJECTIVES: The objectives were (i) to standardize FCM settings for PMP counts on a routine instrument (Cytomics FC500) using size-calibrated fluorescent beads; (ii) to determine intra-instrument and inter-instrument reproducibility; and (iii) to establish PMP values in healthy subjects. METHODS: Using a blend of size-calibrated fluorescent beads (0.5 and 0.9 mum) in a fixed numerical ratio (Megamix), we gated PMPs in a restricted size window. To test intra-instrument and inter-instrument reproducibility, annexin V and CD41 coexpression were used to count PMPs in frozen aliquots of the same platelet-free plasma (PFP) over 4 months and in PFP from 10 healthy subjects on three independent flow cytometers. RESULTS: This calibrated-bead strategy allowed full long-term control of the FCM-based microparticle protocol and reproducible PMP counts over time [coefficient of variation (CV) < 10%]. Optimal settings were easily transferred from one instrument to another, using Megamix as a stable template. Similar PMP counts (CV < 12%) were obtained using the three instruments. With such a standardized FCM protocol, PMP values were established in healthy subjects (n = 60) with significantly higher levels in women than in men [median (1st quartile to 3rd quartile): 1775 microL(-1) (1014-3039 microL(-1)) vs. 656 microL(-1) (407-962 microL(-1))]. CONCLUSIONS: The present strategy provides a new option for PMP count standardization and thus opens the way for multicenter studies. |
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Authors:
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S Robert; P Poncelet; R Lacroix; L Arnaud; L Giraudo; A Hauchard; J Sampol; F Dignat-George |
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Publication Detail:
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Type: Journal Article Date: 2008-10-18 |
Journal Detail:
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Title: Journal of thrombosis and haemostasis : JTH Volume: 7 ISSN: 1538-7836 ISO Abbreviation: J. Thromb. Haemost. Publication Date: 2009 Jan |
Date Detail:
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Created Date: 2008-12-18 Completed Date: 2009-02-27 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101170508 Medline TA: J Thromb Haemost Country: England |
Other Details:
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Languages: eng Pagination: 190-7 Citation Subset: IM |
Affiliation:
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Unité Mixte de Recherche S 608 (UMR-S 608), Institut National de la Santé et de la Recherche Médicale (INSERM), Université de la Méditerranée, Marseille, France. stephane.robert@pharmacie.univ-mrs.fr |
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| MeSH Terms | |
Descriptor/Qualifier:
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Blood Platelets* Calibration Cell-Derived Microparticles / pathology* Flow Cytometry / methods, standards* Humans Microspheres Observer Variation Particle Size Reference Standards |
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