PMID- 30988894 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20200929 IS - 2001-3078 (Print) IS - 2001-3078 (Electronic) IS - 2001-3078 (Linking) VI - 8 IP - 1 DP - 2019 TI - Extracellular vesicle measurements with nanoparticle tracking analysis - An accuracy and repeatability comparison between NanoSight NS300 and ZetaView. PG - 1596016 LID - 10.1080/20013078.2019.1596016 [doi] LID - 1596016 AB - 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 (% D(TEM) range: 79.5-134.3) compared to ZetaView (% D(TEM) 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. FAU - Bachurski, Daniel AU - Bachurski D AUID- ORCID: 0000-0001-9168-9680 AD - Department I of Internal Medicine, University 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 Cologne, Cologne, Germany. FAU - Schuldner, Maximiliane AU - Schuldner M AD - Department I of Internal Medicine, University of Cologne, Cologne, Germany. AD - Experimental Tumor Research, Center for Tumor Biology and Immunology, Department of Hematology, Oncology and Immunology, Philipps University Marburg, Marburg, Germany. FAU - Nguyen, Phuong-Hien AU - Nguyen PH AD - Department I of Internal Medicine, University 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 Cologne, Cologne, Germany. FAU - Malz, Alexandra AU - Malz A AD - Experimental Tumor Research, Center for Tumor Biology and Immunology, Department of Hematology, Oncology and Immunology, Philipps University Marburg, Marburg, Germany. FAU - Reiners, Katrin S AU - Reiners KS AD - Department I of Internal Medicine, University of Cologne, Cologne, Germany. AD - Institute for Clinical Chemistry and Clinical Pharmacology, University of Bonn, Bonn, Germany. FAU - Grenzi, Patricia C AU - Grenzi PC AUID- ORCID: 0000-0001-6494-8288 AD - Department I of Internal Medicine, University of Cologne, Cologne, Germany. FAU - Babatz, Felix AU - Babatz F AD - CECAD Center of Excellence on ''Cellular Stress Responses in Aging-Associated Diseases'', University of Cologne, Cologne, Germany. FAU - Schauss, Astrid C AU - Schauss AC AD - CECAD Center of Excellence on ''Cellular Stress Responses in Aging-Associated Diseases'', University of Cologne, Cologne, Germany. FAU - Hansen, Hinrich P AU - Hansen HP AD - Department I of Internal Medicine, University 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 Cologne, Cologne, Germany. FAU - Hallek, Michael AU - Hallek M AD - Department I of Internal Medicine, University 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 Cologne, Cologne, Germany. FAU - Pogge von Strandmann, Elke AU - Pogge von Strandmann E AD - Experimental Tumor Research, Center for Tumor Biology and Immunology, Department of Hematology, Oncology and Immunology, Philipps University Marburg, Marburg, Germany. LA - eng PT - Journal Article DEP - 20190401 PL - United States TA - J Extracell Vesicles JT - Journal of extracellular vesicles JID - 101610479 PMC - PMC6450530 OTO - NOTNLM OT - Extracellular vesicles OT - accuracy OT - exosomes OT - nanoparticle tracking analysis OT - repeatability OT - reproducibility OT - single particle interferometric reflectance imaging sensing OT - transmission electron microscopy EDAT- 2019/04/17 06:00 MHDA- 2019/04/17 06:01 PMCR- 2019/04/01 CRDT- 2019/04/17 06:00 PHST- 2018/05/18 00:00 [received] PHST- 2019/02/14 00:00 [revised] PHST- 2019/03/03 00:00 [accepted] PHST- 2019/04/17 06:00 [entrez] PHST- 2019/04/17 06:00 [pubmed] PHST- 2019/04/17 06:01 [medline] PHST- 2019/04/01 00:00 [pmc-release] AID - 1596016 [pii] AID - 10.1080/20013078.2019.1596016 [doi] PST - epublish SO - J Extracell Vesicles. 2019 Apr 1;8(1):1596016. doi: 10.1080/20013078.2019.1596016. eCollection 2019.