Characterization of extracellular vesicles and synthetic nanoparticles with four orthogonal single-particle analysis platforms

Autor: Tanina Arab, Barbara S. Slusher, Yiyao Huang, Zezhou Zhao, Norman J. Haughey, Wyatt N. Vreeland, Barbara S. Smith, Olesia Gololobova, Zhaohao Liao, Kenneth J. Pienta, Michael E. Paulaitis, Juan Pablo Tosar, Emily Mallick, Liang Dong, Patrick M. Tarwater, Kenneth W. Witwer, Angela M. Zivkovic
Přispěvatelé: Tosar Rovira Juan Pablo, Universidad de la República (Uruguay). Facultad de Ciencias. Centro de Investigaciones Nucleares., Arab Tanina, Mallick Emily R., Huang Yiyao, Dong Liang, Liao Zhaohao
Rok vydání: 2020
Předmět:
0301 basic medicine
Microfluidics
Analytical chemistry
Nanoparticle
Ultrafiltration
Single particle analysis
nanoflow cytometry
ectosomes
resistive pulse sensing
0302 clinical medicine
Nanotechnology
Research Articles
Microscopy
Chromatography
Gel
education.field_of_study
Chemistry
Fluorescence
Single Molecule Imaging
single particle interferometric reflectance imaging sensing
Transmission electron microscopy
030220 oncology & carcinogenesis
Chromatography
Gel
microvesicles
Research Article
Histology
Size-exclusion chromatography
Population
Nanoparticle tracking analysis
Bioengineering
exosomes
Electron
Cell Line
03 medical and health sciences
Extracellular Vesicles
Microscopy
Electron
Transmission
nanoparticle tracking analysis
Transmission
Humans
Particle Size
education
QH573-671
Cell Biology
030104 developmental biology
Nanoparticles
Polystyrenes
Particle
Biochemistry and Cell Biology
Cytology
Ultracentrifugation
Biomarkers
Zdroj: Journal of Extracellular Vesicles
Journal of Extracellular Vesicles, Vol 10, Iss 6, Pp n/a-n/a (2021)
Journal of extracellular vesicles, vol 10, iss 6
COLIBRI
Universidad de la República
instacron:Universidad de la República
DOI: 10.1101/2020.08.04.237156
Popis: We compared four orthogonal technologies for sizing, counting, and phenotyping of extracellular vesicles (EVs) and synthetic particles. The platforms were: single-particle interferometric reflectance imaging sensing (SP-IRIS) with fluorescence, nanoparticle tracking analysis (NTA) with fluorescence, microfluidic resistive pulse sensing (MRPS), and nanoflow cytometry measurement (NFCM). Results were compared with standard EV characterization techniques such as transmission electron microscopy (TEM) and Western blot (WB). EVs from the human T lymphocyte line H9 (high CD81, low CD63) and the promonocytic line U937 (low CD81, high CD63) were separated from culture conditioned medium (CCM) by differential ultracentrifugation (dUC) or a combination of ultrafiltration (UF) and size exclusion chromatography (SEC) and characterized per MISEV2018 guidelines. Mixtures of synthetic particles (silica and polystyrene spheres) with known sizes and/or concentrations were also tested. MRPS and NFCM returned similar particle counts, while NTA detected counts approximately one order of magnitude lower for EVs, but not for synthetic particles. SP-IRIS events could not be used to estimate particle concentrations. For sizing, SP-IRIS, MRPS, and NFCM returned similar size profiles, with smaller sizes predominating (per power law distribution), but with sensitivity typically dropping off below diameters of 60 nm. NTA detected a population of particles with a mode diameter greater than 100 nm. Additionally, SP-IRIS, MRPS, and NFCM were able to identify at least three of four distinct size populations in a mixture of silica or polystyrene nanoparticles. Finally, for tetraspanin phenotyping, the SP-IRIS platform in fluorescence mode and NFCM were able to detect at least two markers on the same particle. Based on the results of this study, we can draw conclusions about existing single-particle analysis capabilities that may be useful for EV biomarker development and mechanistic studies.
Databáze: OpenAIRE
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