Analysis of Extracellular Vesicles Using Fluorescence Nanoparticle Tracking Analysis

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

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies
  • Cell-Derived Microparticles
  • Exosomes / chemistry
  • Extracellular Vesicles* / chemistry
  • Nanoparticles* / chemistry
  • Quantum Dots
  • Spectrometry, Fluorescence / methods*
  • Trophoblasts / metabolism

Substances

  • Antibodies