Sizing and phenotyping of cellular vesicles using Nanoparticle Tracking Analysis

By R. Dragovic, C. Gardiner, A. Brooks, D. Tannetta, D. Ferguson, P. Hole, B. Carr, Christopher W. G. Redman, Adrian L. Harris, P. Dobson and 2 more. Nanomedicine: Nanotechnology, Biology and Medicine, 2011

1,325 citations recorded by the source corpus.

Cellular microvesicles and nanovesicles (exosomes) are involved in many disease processes and have major potential as biomarkers. However, developments in this area are constrained by limitations in the technology available for their measurement. Here we report on the use of fluorescence nanoparticle tracking analysis (NTA) to rapidly size and phenotype cellular vesicles. In this system vesicles are visualized by light scattering using a light microscope. A video is taken, and the NTA software tracks the brownian motion of individual vesicles and calculates their size and total concentration. Using human placental vesicles and plasma, we have demonstrated that NTA can measure cellular vesicles as small as ∼50 nm and is far more sensitive than conventional flow cytometry (lower limit ∼300 nm). By combining NTA with fluorescence measurement we have demonstrated that vesicles can be labeled with specific antibody-conjugated quantum dots, allowing their phenotype to be determined. From the Clinical Editor The authors of this study utilized fluorescence nanoparticle tracking analysis (NTA) to rapidly size and phenotype cellular vesicles, demonstrating that NTA is far more sensitive than conventional flow cytometry.

Subfields: Cancer Research, Immunology, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/d34f0fe4c256839ae382d9d1222c7e06bab2d5fa

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Cancer Research

1. S. El Andaloussi
2. M. Wood
3. C. Théry
4. P. Vader
5. Jan Lötvall
6. Y. Lee
7. Edit I. Buzás
8. Aled Clayton

In Immunology

1. S. Lim
2. C. Théry
3. S. Pluchino

In Molecular Biology

1. C. Théry
2. M. Wood
3. S. El Andaloussi
4. P. Vader
5. Jan Lötvall
6. Muniyandi Jeyaraj
7. M. Qasim
8. Sangiliyandi Gurunathan

Citations in the corpus

• Extracellular vesicle in vivo biodistribution is determined by cell source, route of administration and targeting (2015, neutral): “NTA determines the size of particles based on Brownian motion and is commonly used for analysis of nanometre-sized particles (29).”
• Biogenesis of extracellular vesicles (EV): exosomes, microvesicles, retrovirus-like vesicles, and apoptotic bodies (2013, neutral): “The purity of the preparation is typically confirmed using electron microscopy [19] or laser scatter tracking [21].”
• How pure are your vesicles? (2013, result): “94 nm for the averaged histogram, which sits within the expected range of size discrimination of the instrument, and compares with other NanoSight-based observations (9).”
• Biogenesis, secretion, and intercellular interactions of exosomes and other extracellular vesicles. (2014, neutral): “A device allowing Nanoparticle Tracking Analysis (NTA) has been designed to measure the size distribution and concentration of nanoparticles (Dragovic et al. 2011).”
• Review of the Isolation, Characterization, Biological Function, and Multifarious Therapeutic Approaches of Exosomes (2019, methodology): “Therefore, ultracentrifugation remains one of the most reliable techniques, followed by western blotting, NTA [12], and electron microscopy.”
• Hypoxic enhancement of exosome release by breast cancer cells (2012, methodology): “Isolated exosomes were analysed using the Nanosight LM10 system (Nanosight Ltd) [29,30] equipped with a blue laser (405 nm).”
• Applying extracellular vesicles based therapeutics in clinical trials – an ISEV position paper (2015, background): “Although NTA is most frequently used, all these technologies have their specific pitfalls (320).”
• Standardization of sample collection, isolation and analysis methods in extracellular vesicle research (2013, neutral): “Although standard light scatter NTA does not detect biochemical composition or cellular origin of vesicles, analysis of fluorescently labelled vesicles is also feasible (256,260).”
• Exosomes and microvesicles: extracellular vesicles for genetic information transfer and gene therapy. (2012, methodology): “In addition, new technologies such as the Nanosight (101), new FACS (102) protocols and curation of an exosome marker database like exocarta.”