Microparticles (MPs) are potential noninvasive biomarkers for diagnosis or prognosis
in pathologic conditions. However, the lack of standardization of the preanalytical
and analytical methods leads to a wide variability in MPs results. The recently developed
Megamix-Plus beads, a new bead-based standardization tool optimized to specific types
of flow cytometers, could help circumvent this problem. The aim of the present study
was to determine whether the number of total MPs and platelet-derived MPs (PMPs) is
similar using 2 different cytometer platforms calibrated with the Megamix-Plus beads.
Blood samples from 65 patients with deep venous thrombosis were collected and processed
to obtain platelet poor plasma (PPP). The number of total MPs and PMPs in each PPP
sample was measured using 2 flow cytometers. Megamix-Plus side scatter channel beads
were used to calibrate the LSRFortessa flow cytometer from Becton Dickinson, whereas
Megamix-Plus forward scatter channel beads were applied to the Navios flow cytometer
from Beckman Coulter. High correlation of total MPs and PMPs values between the flow
cytometers was found (r = 0.908, P < 0.01 and r = 0.910, P < 0.001, respectively). However, the absolute numbers of total MPs and PMPs were
significantly higher measured with the Navios flow cytometer compared with the LSRFortessa
cytometer. Therefore, both platforms are valid for MPs determination in general, although
a similar platform with the same calibration tool could be a better choice for multicenter
studies.
Abbreviations:
MPs (microparticles), PPP (platelet poor plasma), PMPs (platelet MPs), BC (Beckman Coulter), BD (Becton Dickinson), FS/FSC (forward Scatter/channel), SS/SSC (side Scatter/channel), PE (phycoerythrin), PECy7 (phycoerythrin-Cyanine7), FITC (fluorescein isothiocyanate), FL1 (fluorescence 1 channel)To read this article in full you will need to make a payment
Purchase one-time access:
Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online accessOne-time access price info
- For academic or personal research use, select 'Academic and Personal'
- For corporate R&D use, select 'Corporate R&D Professionals'
Subscribe:
Subscribe to Translational ResearchAlready a print subscriber? Claim online access
Already an online subscriber? Sign in
Register: Create an account
Institutional Access: Sign in to ScienceDirect
References
- Formation of procoagulant microparticles and properties.Thromb Res. 2010; 125: S46-S48
- Microparticles: major transport vehicles for distinct microRNAs in circulation.Cardiovasc Res. 2012; 93: 633-644
- Microparticles, malignancy and thrombosis.Br J Haematol. 2011; 152: 688-700
- Endothelial-derived microparticles: biological conveyors at the crossroad of inflammation, thrombosis and angiogenesis.Thromb Haemost. 2010; 104: 456-463
- Mechanisms of thrombus formation.N Engl J Med. 2008; 359: 938-949
- Microparticles: protagonists of a novel communication network for intercellular information exchange.Circ Res. 2010; 107: 1047-1057
- Circulating endothelial cells and procoagulant microparticles in patients with glioblastoma: prognostic value.PLoS One. 2013; 8: e69034
- Association of circulating endothelial microparticles with cardiometabolic risk factors in the Framingham Heart Study.Eur Heart J. 2014; 35: 2972-2979
- Altered profile of circulating microparticles in rheumatoid arthritis patients.Clin Sci (Lond). 2015; 128: 437-448
- Impact of pre-analytical parameters on the measurement of circulating microparticles: towards standardization of protocol.J Thromb Haemost. 2012; 10: 437-446
- Measurement of circulating cell-derived microparticles by flow cytometry: sources of variability within the assay.Thromb Res. 2011; 127: 370-377
- Flow cytometric detection of endothelial microparticles (EMP): effects of centrifugation and storage alter with the phenotype studied.Thromb Res. 2010; 125: 332-339
- Standardization of pre-analytical variables in plasma microparticle determination: results of the International Society on Thrombosis and Haemostasis SSC Collaborative workshop.J Thromb Haemost. 2013; 11: 1190-1193
- Pre-analytical issues in the measurement of circulating microparticles: current recommendations and pending questions.J Thromb Haemost. 2013; 11: 693-696
- High-sensitivity flow cytometry provides access to standardized measurement of small-size microparticles–brief report.Arterioscler Thromb Vasc Biol. 2012; 32: 1054-1058
- Recovery and composition of microparticles after snap-freezing depends on thawing temperature.Blood Coagul Fibrinolysis. 2009; 20: 52-56
- Single vs. swarm detection of microparticles and exosomes by flow cytometry.J Thromb Haemost. 2012; 10: 919-930
- A new microparticle size calibration standard for use in measuring smaller microparticles using a new flow cytometer.J Thromb Haemost. 2011; 9: 1216-1224
- Silica microspheres are superior to polystyrene for microvesicle analysis by flow cytometry.Thromb Res. 2015; 135: 1000-1006
- Standardization of platelet-derived microparticle counting using calibrated beads and a Cytomics FC500 routine flow cytometer: a first step towards multicenter studies?.J Thromb Haemost. 2009; 7: 190-197
- Standardization of platelet-derived microparticle enumeration by flow cytometry with calibrated beads: results of the International Society on Thrombosis and Haemostasis SSC Collaborative workshop.J Thromb Haemost. 2010; 8: 2571-2574
- Standardization of flow cytometry-based microparticle analysis for cytometers using side scatter for size measurements.J Thromb Haemost. 2012; 11: FVB20
- Standardized counting of circulating platelet microparticles using currently available flow cytometers and scatter-based triggering: forward or side scatter?.Cytometry A. 2015; ([Epub ahead of print])https://doi.org/10.1002/cyto.a.22685
Article info
Publication history
Published online: August 20, 2015
Accepted:
August 13,
2015
Received in revised form:
August 4,
2015
Received:
April 13,
2015
Identification
Copyright
© 2015 Elsevier Inc. Published by Elsevier Inc. All rights reserved.
