Universität zu Köln

Bachurski, Daniel ORCID: 0000-0001-9168-9680, Schuldner, Maximiliane, Nguyen, Phuong-Hien, Malz, Alexandra, Reiners, Katrin S., Grenzi, Patricia C., Babatz, Felix, Schauss, Astrid C., Hansen, Hinrich P., Hallek, Michael and von Strandmann, Elke Pogge (2019). Extracellular vesicle measurements with nanoparticle tracking analysis - An accuracy and repeatability comparison between NanoSight NS300 and ZetaView. J. Extracell. Vesicles, 8 (1). ABINGDON: TAYLOR & FRANCIS LTD. ISSN 2001-3078

Full text not available from this repository.

Abstract

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.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Bachurski, Daniel UNSPECIFIED orcid.org/0000-0001-9168-9680 UNSPECIFIED Schuldner, Maximiliane UNSPECIFIED UNSPECIFIED UNSPECIFIED Nguyen, Phuong-Hien UNSPECIFIED UNSPECIFIED UNSPECIFIED Malz, Alexandra UNSPECIFIED UNSPECIFIED UNSPECIFIED Reiners, Katrin S. UNSPECIFIED UNSPECIFIED UNSPECIFIED Grenzi, Patricia C. UNSPECIFIED UNSPECIFIED UNSPECIFIED Babatz, Felix UNSPECIFIED UNSPECIFIED UNSPECIFIED Schauss, Astrid C. UNSPECIFIED UNSPECIFIED UNSPECIFIED Hansen, Hinrich P. UNSPECIFIED UNSPECIFIED UNSPECIFIED Hallek, Michael UNSPECIFIED UNSPECIFIED UNSPECIFIED von Strandmann, Elke Pogge UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-139225
DOI:	10.1080/20013078.2019.1596016
Journal or Publication Title:	J. Extracell. Vesicles
Volume:	8
Number:	1
Date:	2019
Publisher:	TAYLOR & FRANCIS LTD
Place of Publication:	ABINGDON
ISSN:	2001-3078
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language PARTICLE-SIZE; QUANTIFICATION; EXOSOMES Multiple languages Cell Biology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/13922