Extracellular vesicle measurements with nanoparticle tracking analysis – An accuracy and repeatability comparison between NanoSight NS300 and ZetaView
The expanding field of extracellular vesicle (EV) research needs reproducible and accurate methods to characterize single EVs. Nanoparticle Tracking Analysis (NTA) is commonly used to determine EV concentration and diameter. As the EV field is lacking methods to easily confirm and validate NTA data,...
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Online Access: | http://dx.doi.org/10.1080/20013078.2019.1596016 |
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doaj-37bb946283c0421493d4c62f7aabbed82020-11-25T01:34:23ZengTaylor & Francis GroupJournal of Extracellular Vesicles2001-30782019-12-018110.1080/20013078.2019.15960161596016Extracellular vesicle measurements with nanoparticle tracking analysis – An accuracy and repeatability comparison between NanoSight NS300 and ZetaViewDaniel Bachurski0Maximiliane Schuldner1Phuong-Hien Nguyen2Alexandra Malz3Katrin S Reiners4Patricia C Grenzi5Felix Babatz6Astrid C Schauss7Hinrich P Hansen8Michael Hallek9Elke Pogge von Strandmann10University Hospital of Cologne, Center for Integrated Oncology Cologne-Bonn, CECAD Center of Excellence on ‘‘Cellular Stress Responses in Aging-Associated Diseases’’, Center for Molecular Medicine Cologne, University of CologneUniversity of CologneUniversity Hospital of Cologne, Center for Integrated Oncology Cologne-Bonn, CECAD Center of Excellence on ‘‘Cellular Stress Responses in Aging-Associated Diseases’’, Center for Molecular Medicine Cologne, University of CologneOncology and Immunology, Philipps University MarburgUniversity of CologneUniversity of CologneUniversity of CologneUniversity of CologneUniversity Hospital of Cologne, Center for Integrated Oncology Cologne-Bonn, CECAD Center of Excellence on ‘‘Cellular Stress Responses in Aging-Associated Diseases’’, Center for Molecular Medicine Cologne, University of CologneUniversity Hospital of Cologne, Center for Integrated Oncology Cologne-Bonn, CECAD Center of Excellence on ‘‘Cellular Stress Responses in Aging-Associated Diseases’’, Center for Molecular Medicine Cologne, University of CologneOncology and Immunology, Philipps University MarburgThe expanding field of extracellular vesicle (EV) research needs reproducible and accurate methods to characterize single EVs. Nanoparticle Tracking Analysis (NTA) is commonly used to determine EV concentration and diameter. As the EV field is lacking methods to easily confirm and validate NTA data, questioning the reliability of measurements remains highly important. In this regard, a comparison addressing measurement quality between different NTA devices such as Malvern’s NanoSight NS300 or Particle Metrix’ ZetaView has not yet been conducted. To evaluate the accuracy and repeatability of size and concentration determinations of both devices, we employed comparative methods including transmission electron microscopy (TEM) and single particle interferometric reflectance imaging sensing (SP-IRIS) by ExoView. Multiple test measurements with nanospheres, liposomes and ultracentrifuged EVs from human serum and cell culture supernatant were performed. Additionally, serial dilutions and freeze-thaw cycle-dependent EV decrease were measured to determine the robustness of each system. Strikingly, NanoSight NS300 exhibited a 2.0–2.1-fold overestimation of polystyrene and silica nanosphere concentration. By measuring serial dilutions of EV samples, we demonstrated higher accuracy in concentration determination by ZetaView (% BIAS range: 2.7–8.5) in comparison with NanoSight NS300 (% BIAS range: 32.9–36.8). The concentration measurements by ZetaView were also more precise (% CV range: 0.0–4.7) than measurements by NanoSight NS300 (% CV range: 5.4–10.7). On the contrary, quantitative TEM imaging indicated more accurate EV sizing by NanoSight NS300 (% DTEM range: 79.5–134.3) compared to ZetaView (% DTEM range: 111.8–205.7), while being equally repeatable (NanoSight NS300% CV range: 0.8–6.7; ZetaView: 1.4–7.8). However, both devices failed to report a peak EV diameter below 60 nm compared to TEM and SP-IRIS. Taken together, NTA devices differ strongly in their hardware and software affecting measuring results. ZetaView provided a more accurate and repeatable depiction of EV concentration, whereas NanoSight NS300 supplied size measurements of higher resolution.http://dx.doi.org/10.1080/20013078.2019.1596016extracellular vesiclesexosomesnanoparticle tracking analysistransmission electron microscopysingle particle interferometric reflectance imaging sensingaccuracyrepeatabilityreproducibility |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Daniel Bachurski Maximiliane Schuldner Phuong-Hien Nguyen Alexandra Malz Katrin S Reiners Patricia C Grenzi Felix Babatz Astrid C Schauss Hinrich P Hansen Michael Hallek Elke Pogge von Strandmann |
spellingShingle |
Daniel Bachurski Maximiliane Schuldner Phuong-Hien Nguyen Alexandra Malz Katrin S Reiners Patricia C Grenzi Felix Babatz Astrid C Schauss Hinrich P Hansen Michael Hallek Elke Pogge von Strandmann Extracellular vesicle measurements with nanoparticle tracking analysis – An accuracy and repeatability comparison between NanoSight NS300 and ZetaView Journal of Extracellular Vesicles extracellular vesicles exosomes nanoparticle tracking analysis transmission electron microscopy single particle interferometric reflectance imaging sensing accuracy repeatability reproducibility |
author_facet |
Daniel Bachurski Maximiliane Schuldner Phuong-Hien Nguyen Alexandra Malz Katrin S Reiners Patricia C Grenzi Felix Babatz Astrid C Schauss Hinrich P Hansen Michael Hallek Elke Pogge von Strandmann |
author_sort |
Daniel Bachurski |
title |
Extracellular vesicle measurements with nanoparticle tracking analysis – An accuracy and repeatability comparison between NanoSight NS300 and ZetaView |
title_short |
Extracellular vesicle measurements with nanoparticle tracking analysis – An accuracy and repeatability comparison between NanoSight NS300 and ZetaView |
title_full |
Extracellular vesicle measurements with nanoparticle tracking analysis – An accuracy and repeatability comparison between NanoSight NS300 and ZetaView |
title_fullStr |
Extracellular vesicle measurements with nanoparticle tracking analysis – An accuracy and repeatability comparison between NanoSight NS300 and ZetaView |
title_full_unstemmed |
Extracellular vesicle measurements with nanoparticle tracking analysis – An accuracy and repeatability comparison between NanoSight NS300 and ZetaView |
title_sort |
extracellular vesicle measurements with nanoparticle tracking analysis – an accuracy and repeatability comparison between nanosight ns300 and zetaview |
publisher |
Taylor & Francis Group |
series |
Journal of Extracellular Vesicles |
issn |
2001-3078 |
publishDate |
2019-12-01 |
description |
The expanding field of extracellular vesicle (EV) research needs reproducible and accurate methods to characterize single EVs. Nanoparticle Tracking Analysis (NTA) is commonly used to determine EV concentration and diameter. As the EV field is lacking methods to easily confirm and validate NTA data, questioning the reliability of measurements remains highly important. In this regard, a comparison addressing measurement quality between different NTA devices such as Malvern’s NanoSight NS300 or Particle Metrix’ ZetaView has not yet been conducted. To evaluate the accuracy and repeatability of size and concentration determinations of both devices, we employed comparative methods including transmission electron microscopy (TEM) and single particle interferometric reflectance imaging sensing (SP-IRIS) by ExoView. Multiple test measurements with nanospheres, liposomes and ultracentrifuged EVs from human serum and cell culture supernatant were performed. Additionally, serial dilutions and freeze-thaw cycle-dependent EV decrease were measured to determine the robustness of each system. Strikingly, NanoSight NS300 exhibited a 2.0–2.1-fold overestimation of polystyrene and silica nanosphere concentration. By measuring serial dilutions of EV samples, we demonstrated higher accuracy in concentration determination by ZetaView (% BIAS range: 2.7–8.5) in comparison with NanoSight NS300 (% BIAS range: 32.9–36.8). The concentration measurements by ZetaView were also more precise (% CV range: 0.0–4.7) than measurements by NanoSight NS300 (% CV range: 5.4–10.7). On the contrary, quantitative TEM imaging indicated more accurate EV sizing by NanoSight NS300 (% DTEM range: 79.5–134.3) compared to ZetaView (% DTEM range: 111.8–205.7), while being equally repeatable (NanoSight NS300% CV range: 0.8–6.7; ZetaView: 1.4–7.8). However, both devices failed to report a peak EV diameter below 60 nm compared to TEM and SP-IRIS. Taken together, NTA devices differ strongly in their hardware and software affecting measuring results. ZetaView provided a more accurate and repeatable depiction of EV concentration, whereas NanoSight NS300 supplied size measurements of higher resolution. |
topic |
extracellular vesicles exosomes nanoparticle tracking analysis transmission electron microscopy single particle interferometric reflectance imaging sensing accuracy repeatability reproducibility |
url |
http://dx.doi.org/10.1080/20013078.2019.1596016 |
work_keys_str_mv |
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