2018
DOI: 10.1080/20013078.2018.1454776
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Improved resolution in extracellular vesicle populations using 405 instead of 488 nm side scatter

Abstract: Improvements in identification and assessment of extracellular vesicles (EVs) have fuelled a recent surge in EV publications investigating their roles as biomarkers and mediators of disease. Meaningful scientific comparisons are, however, hampered by difficulties in accurate, reproducible enumeration and characterization of EVs in biological fluids. High-sensitivity flow cytometry (FCM) is presently the most commonly applied strategy to assess EVs, yet its utility is limited by variant ability to resolve small… Show more

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Cited by 49 publications
(38 citation statements)
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“…Particle number can be measured by light scattering technologies, such as nanoparticle tracking analysis (NTA); by standard flow cytometry for larger EVs [164][165][166][167] or high resolution flow cytometry for smaller EVs [127,[168][169][170][171][172][173][174][175][176]; by resistive pulse sensing (RPS) for a wide range of sizes, depending on pore size [177]; by cryo-EM [174]; by a platform combining surface plasmon resonance (SPR) with AFM [178]; or by other techniques with similar capabilities. Accurate quantitation may be possible only within a certain concentration and size range that varies by platform; where possible, this range (or the minimum and maximum diameter measured) should be reported along with concentration.…”
Section: ) Intermediate Recovery Intermediate Specificity: Methods mentioning
confidence: 99%
“…Particle number can be measured by light scattering technologies, such as nanoparticle tracking analysis (NTA); by standard flow cytometry for larger EVs [164][165][166][167] or high resolution flow cytometry for smaller EVs [127,[168][169][170][171][172][173][174][175][176]; by resistive pulse sensing (RPS) for a wide range of sizes, depending on pore size [177]; by cryo-EM [174]; by a platform combining surface plasmon resonance (SPR) with AFM [178]; or by other techniques with similar capabilities. Accurate quantitation may be possible only within a certain concentration and size range that varies by platform; where possible, this range (or the minimum and maximum diameter measured) should be reported along with concentration.…”
Section: ) Intermediate Recovery Intermediate Specificity: Methods mentioning
confidence: 99%
“…Because the diameter of the MPs is too smaller to detect with conventional method (488‐nm SSC laser light). So in order to improve the sensitivity for MP detection, we adjusted the side scatter (SSC) with 405‐nm (instead of 488 nm) laser light to avoid background noise during acquisition . The predefined MP gate was applied to all samples during analysis.…”
Section: Methodsmentioning
confidence: 99%
“…Lower EV detection limits of 45 nm with NTA and 100 nm by flow virometry were reported in this work. The sample composition and also the implementation of the V-SSC instead of the 488 nm SSC in flow virometry could contribute to explain these differences [34]. Still, a higher level of accuracy is possible by using National Institute of Standards and Technology (NIST)-traceable beads in the Mie correlation with flow virometry.…”
Section: Particle Size Distributionmentioning
confidence: 99%
“…Still, a significant loss of scattered light that fall in the range of the background noise of the instrument and different sensitivities between equipments are a general concern [33]. To address this issue, the implementation of the violet (405 nm) side scatter (V-SSC) has been reported to improve the sensitivity but also the resolution of the technique [34]. Owing to the specific features of each analytical method, characterization results such as particle concentration obtained by different techniques are often difficult to compare.…”
Section: Introductionmentioning
confidence: 99%