Novel test for microparticles in platelet‐rich plasma and platelet concentrates using dynamic light scattering

Xu, Yan; Nakane, Nobu; Maurer‐Spurej, Elisabeth

doi:10.1111/j.1537-2995.2010.02819.x

Cited by 54 publications

(39 citation statements)

References 18 publications

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“…As previously shown by electron microscopy, 14 new techniques such as laser-induced nanotracking, 16,17 atomic force microscopy, 18,19 and dynamic light scattering [20][21][22] recently confirmed that the major part of MP counts falls below the detection threshold of sd-FCM. However, recent FCM innovations strive for an MP numeration closer to these sensitive techniques while giving supplemental information such as immuno-phenotype.…”

Section: Discussionmentioning

confidence: 76%

High-Sensitivity Flow Cytometry Provides Access to Standardized Measurement of Small-Size Microparticles—Brief Report

Robert

Lacroix²,

Poncelet³

et al. 2012

ATVB

142

137

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Objective-Cellular microparticles (MP) are promising biomarkers in many pathological situations. Although flow cytometry (FCM) is widely used for their measurement, it has raised controversies because the smallest MP size falls below the detection limit of standard FCM (sd-FCM). Following recent technological improvements leading to high sensitivity FCM (hs-FCM), our objectives were (1) to evaluate the potential of hs-FCM for extended MP detection, (2) to set up a standardized protocol for MP enumeration, and (3) to compare MP counts obtained with both sensitivity levels. Methods and Results-Compared with sd-FCM, hs-FCM displayed improved forward scatter resolution and lower background noise, allowing us to discriminate previously undetectable small MP in plasma samples. Using fluorescent beads with appropriate sizes (0.1/0.3/0.5/0.9 m) and relative amounts, a new standardized hs-FCM MP protocol was set up and provided reproducible MP counts. Applied to coronary patient samples, it resulted into 8-to 20-fold increases in MP counts as compared with sd-FCM. Interestingly, the ratio between small and large MP varied according to clinical status but also depending on MP subset, suggesting access to new biological information. 3-5 Circulating MP are elevated in many prothrombotic and inflammatory disorders; cardiovascular, autoimmune, and infectious diseases; and cancer. 4,[6][7][8][9][10] Although MP counts may provide useful diagnostic/prognostic information, assessment of their pathophysiological relevance is hampered by methodological concerns. Flow cytometry (FCM) is the most commonly used technique, 11 allowing both enumeration and characterization of MP cellular origin with high throughput. The forward scatter (FS) parameter is generally used to define the analysis gate for MP. Unfortunately, because of their limited FS sensitivity, standard flow cytometers measure only a small part of the MP population. Recently, high-sensitivity flow cytometers with significantly improved light scatter detection became available. As we first reported, these instruments resolve a previously undetectable subpopulation of small-size MPs. Conclusion-Recent12 Among the technical improvements found in high sensitivity FCM (hs-FCM), a new option of the Beckman Coulter Gallios called W2 increases FS resolution by amplifying the FS signals collected at large angles. 13 Whether this option can be used in a standardized manner, to improve the detection of MPs derived from platelets (PMP), erythrocytes (Ery-MP), leukocytes (Leu-MP), and endothelial cells (EMP), remains an open question. Our objectives were then (1) to evaluate the impact of high-sensitivity flow cytometer technical improvement on the detection of small-size MP, (2) to set up a new standardized protocol based on calibrated microbeads for MP enumeration, and (3) to compare MP counts obtained at both sensitivity levels of FCM. MethodsBlood and plasma samples, MP preparation and flow cytometric analysis, size-conditioned MP filtration, intra-and inter-instrument reproducibi...

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Section: Discussionmentioning

confidence: 76%

High-Sensitivity Flow Cytometry Provides Access to Standardized Measurement of Small-Size Microparticles—Brief Report

Robert

Lacroix²,

Poncelet³

et al. 2012

ATVB

142

137

View full text Add to dashboard Cite

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“…For example, it is expected that size distributions of polydisperse samples are biased toward small numbers of large particles [44,45], such as platelets and other contaminants, because such particles scatter light more efficiently than small vesicles. Therefore, DLS typically obtains a more than twofold increase in the median diameter of vesicles from plasma compared with other novel techniques [6,12,16,46,47]. We conclude that DLS requires careful data interpretation and may be a useful method provided that the shape of the size distribution is known.…”

Section: Dynamic Light Scatteringmentioning

confidence: 83%

Innovation in detection of microparticles and exosomes

Pol

Coumans

Varga

et al. 2013

Journal of Thrombosis and Haemostasis

226

194

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Summary. Cell-derived or extracellular vesicles, including microparticles and exosomes, are abundantly present in body fluids such as blood. Although such vesicles have gained strong clinical and scientific interest, their detection is difficult because many vesicles are extremely small with a diameter of less than 100 nm, and, moreover, these vesicles have a low refractive index and are heterogeneous in both size and composition. In this review, we focus on the relatively high throughput detection of vesicles in suspension by flow cytometry, resistive pulse sensing, and nanoparticle tracking analysis, and we will discuss their applicability and limitations. Finally, we discuss four methods that are not commercially available: Raman microspectroscopy, micro nuclear magnetic resonance, small-angle X-ray scattering (SAXS), and anomalous SAXS. These methods are currently being explored to study vesicles and are likely to offer novel information for future developments.

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“…The microparticles are gated in the forward versus side scatter dot plot. In flow cytometry, the concentration of microparticles is determined relative to a known concentration of fluorescently labeled microspheres with a size of 1,000 nm as previously described [20]. In apheresis platelets the concentration of microparticles calculated from the ThromboLUX score strongly correlated with the microparticle content determined by flow cytometry ( fig.…”

Section: Comparison Of Thrombolux Results With Flow Cytometrymentioning

confidence: 99%

“…Both platelet viability and the number of microparticles are very challenging to assess by microscopy. Microparticles are extremely difficult to capture due to their small diameter of about 200-400 nm and their quick movement in and out of focus [20]. Furthermore, microscopy scoring methods such as the modified Kunicki morphology score ignore potentially important characteristics of platelet concentrates such as microparticle content, microaggregates, and other unclassified particles [3].…”

Section: Comparison Of Thrombolux Results To Microscopymentioning

confidence: 99%

Characterization of Platelet Concentrates Using Dynamic Light Scattering

Labrie¹,

Marshall

Bedi

et al. 2013

Transfus Med Hemother

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Background: Each year, millions of platelet transfusions save the lives of cancer patients and patients with bleeding complications. However, between 10 and 30% of all platelet transfusions are clinically ineffective as measured by corrected count increments, but no test is currently used to identify and avoid these transfusions. ThromboLUX® is the first platelet test intended to routinely characterize platelet concentrates prior to transfusion. Methods: ThromboLUX is a non-invasive, optical test utilizing dynamic light scattering to characterize a platelet sample by the relative quantity of platelets, microparticles, and other particles present in the sample. ThromboLUX also determines the response of platelets to temperature changes. From this information the ThromboLUX score is calculated. Increasing scores indicate increasing numbers of discoid platelets and fewer microparticles. ThromboLUX uses calibrated polystyrene beads as a quality control standard, and accurately measures the size of the beads at multiple temperatures. Results: Results from apheresis concentrates showed that ThromboLUX can determine the microparticle content in unmodified samples of platelet concentrates which correlates well with the enumeration by flow cytometry. ThromboLUX detection of microparticles and microaggregates was confirmed by microscopy. Conclusion: ThromboLUX provides a comprehensive and novel analysis of platelet samples and has potential as a non-invasive routine test to characterize platelet products to identify and prevent ineffective transfusions.

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Novel test for microparticles in platelet‐rich plasma and platelet concentrates using dynamic light scattering

Cited by 54 publications

References 18 publications

High-Sensitivity Flow Cytometry Provides Access to Standardized Measurement of Small-Size Microparticles—Brief Report

High-Sensitivity Flow Cytometry Provides Access to Standardized Measurement of Small-Size Microparticles—Brief Report

Innovation in detection of microparticles and exosomes

Characterization of Platelet Concentrates Using Dynamic Light Scattering

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