Analysis of extracellular vesicles using fluorescence nanoparticle tracking analysis

Carnell-Morris, P., Tannetta, D., Siupa, A., Hole, P. and Dragovic, R. (2017) Analysis of extracellular vesicles using fluorescence nanoparticle tracking analysis. In: Extracellular Vesicles. Springer, pp. 153-173. ISBN 9781493972531 doi: 10.1007/978-1-4939-7253-1_13

Abstract/Summary

Fluorescence nanoparticle tracking analysis (fl-NTA) allows for accurate sizing, counting, and phenotyping of extracellular vesicles (EV). Here, we present two protocols for the analysis of EVs using fl-NTA, highlighting the potential pitfalls and challenges. The first protocol utilizes CellMask Orange™ (CMO) as a general membrane marker to label EVs derived from plasma. The second protocol describes the use of a Qdot-conjugated antibody to identify syncytiotrophoblast (STB)-derived EVs. "Standard" preparations of STB-derived EVs enriched for either microvesicles (STBMV) or exosomes (STBEX), containing a known amount of EV positive for the STB specific antigen placental alkaline phosphatase (PLAP), were also used to optimize fl-NTA camera settings.