2020
DOI: 10.1002/cyto.a.24029
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Fluorescence and Light Scatter Calibration Allow Comparisons of Small Particle Data in Standard Units across Different Flow Cytometry Platforms and Detector Settings

Abstract: Flow cytometers have been utilized for the analysis of submicron‐sized particles since the late 1970s. Initially, virus analyses preceded extracellular vesicle (EV), which began in the 1990s. Despite decades of documented use, the lack of standardization in data reporting has resulted in a growing body of literature that cannot be easily interpreted, validated, or reproduced. This has made it difficult for objective assessments of both assays and instruments, in‐turn leading to significant hindrances in scient… Show more

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Cited by 52 publications
(67 citation statements)
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“…This is also the case for other techniques where affinity-based phenotyping is not or cannot be done. Currently, only single-EV flow cytometry combines abilities for sizing, concentration measurement, affinity-based phenotyping and high throughput with calibration into standard units to provide a limit of sensitivity for each parameter [26][27][28][29]. While not all techniques have a discernable limit of sensitivity that can be derived from a calibration and expressed in standard units, it is possible to perform indirect assessments of sensitivity using reference materials.…”
Section: Current Techniques: An Overviewmentioning
confidence: 99%
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“…This is also the case for other techniques where affinity-based phenotyping is not or cannot be done. Currently, only single-EV flow cytometry combines abilities for sizing, concentration measurement, affinity-based phenotyping and high throughput with calibration into standard units to provide a limit of sensitivity for each parameter [26][27][28][29]. While not all techniques have a discernable limit of sensitivity that can be derived from a calibration and expressed in standard units, it is possible to perform indirect assessments of sensitivity using reference materials.…”
Section: Current Techniques: An Overviewmentioning
confidence: 99%
“…Ideally, the calibration reference materials would also be certified, so as to give a traceable measurement back to SI units and limiting the potential for bias in calibration accuracy. The development of certified calibration reference particles is crucial for instrument measurement calibration, instrument sensitivity quantification and in turn instrumentation standardization and comparisons [29]. Calibration reference materials allow standardized reporting and consequently, validation of published EV studies between assays, instruments and laboratories.…”
Section: Calibrationmentioning
confidence: 99%
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“…The lack of clarity within the nomenclature is in part due to the lack of an ability to separate and characterize all EVs. Today, as flow cytometers become ever more sensitive due to technological advancements, the ability to characterize particles of 100 nm in diameter or smaller is feasible using some commercially available instruments (de Rond et al, 2019;Gasecka et al, 2020;Morales-Kastresana et al, 2019;Stoner et al, 2016;Tian et al, 2020;Tian et al, 2018;Welsh, Jones, & Tang, 2020;Zhu et al, 2014). As the use of small particle flow cytometry increases, it is critical for researchers to understand the particular controls and considerations that are distinctively necessary in small particle flow cytometry, as compared to what is necessary in cellular flow cytometry (see Current Protocols article; Nolan, 2015).…”
Section: Commentary Background Informationmentioning
confidence: 99%