Extracellular vesicle sizing and enumeration by nanoparticle tracking analysis - PubMed (original) (raw)

Extracellular vesicle sizing and enumeration by nanoparticle tracking analysis

Chris Gardiner et al. J Extracell Vesicles. 2013.

Abstract

Nanoparticle tracking analysis (NTA) is a light-scattering technique that is useful for the rapid sizing and enumeration of extracellular vesicles (EVs). As a relatively new method, NTA has been criticised for a lack of standardisation. We propose the use of silica microspheres for the calibration of NTA measurements and describe in detail a protocol for the analysis of EVs by NTA which should minimise many of the sources of variability and imprecision associated with this technique.

Keywords: extracellular vesicles; light scattering; nanoparticle tracking analysis; standardisation.

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Figures

Fig. 1

Fig. 1

Onscreen images showing (A) the correct position of the “thumbprint” at the zero position; (B) overexposed particles due to inappropriately high camera settings; (C) a correctly focussed image of an appropriate concentration of particles; poorly focussed particles due to the stage being (D) too low or (E) too high; and a sample that is too concentrated for analysis.

Fig. 2

Fig. 2

(A) The effect of minimum track length (MTL) on measured size distribution of monodisperse 100 nm silica microspheres; (B) the effect of using automatic (Auto) or manual (MTL5) minimum track length on measurement of a low concentration of polydisperse EVs; (C) the effect of increasing camera level (level 3 to level 7) on the measurement of a mixture of 100 nm and 200 nm microspheres (concentration 10×108/ml and 0.5×108/ml, respectively); and (D) NTA analysis of plasma EVs labelled with CellMask using light scattering (Scatter) and fluorescence (Fluor) measurement.

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References

    1. Dragovic RA, Gardiner C, Brooks AS, Tannetta DS, Ferguson DJ, Hole P, et al. Sizing and phenotyping of cellular vesicles using nanoparticle tracking analysis. Nanomedicine. 2011;7:780–8. - PMC - PubMed
    1. Saveyn H, De BB, Thas O, Hole P, Smith J, Van der Meeren P. Accurate particle size distribution determination by nanoparticle tracking analysis based on 2-D Brownian dynamics simulation. J Colloid Interface Sci. 2010;352:593–600. - PubMed
    1. Van der Meeren P, Kasinos M, Saveyn H. Relevance of two-dimensional Brownian motion dynamics in applying nanoparticle tracking analysis. Methods Mol Biol. 2012;906:525–34. - PubMed
    1. Carr R, Hole P, Malloy A, Nelson P, Smith J. Applications of nanoparticle tracking analysis (NTA) in nanoparticle research – a mini-review. Eur J Parent Pharmaceut Sci. 2009;14:45–50.
    1. Filipe V, Hawe A, Jiskoot W. Critical evaluation of nanoparticle tracking analysis (NTA) by NanoSight for the measurement of nanoparticles and protein aggregates. Pharm Res. 2010;27:796–810. - PMC - PubMed

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