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. 2017:1660:153-173.
doi: 10.1007/978-1-4939-7253-1_13.

Analysis of Extracellular Vesicles Using Fluorescence Nanoparticle Tracking Analysis

Pauline Carnell-Morris  1 , Dionne Tannetta  2 , Agnieszka Siupa  1 , Patrick Hole  1 , Rebecca Dragovic  3
Affiliations

Analysis of Extracellular Vesicles Using Fluorescence Nanoparticle Tracking Analysis

Pauline Carnell-Morris et al. Methods Mol Biol. 2017.

Abstract

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.

Keywords: Exosomes; Extracellular vesicles; Fluorescence nanoparticle tracking analysis; Microvesicles; Quantum dots.

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