Characterizing Extracellular Vesicles Using Nanoparticle-Tracking Analysis

Extracellular vesicles (EVs) are nanosized lipid bound particles secreted by cells. EVs transfer biologic material and can be found in all body fluids. Accurately characterizing the size and concentration of EVs is difficult because of the nanoscale size of EVs. An evolving solution to this problem is tracking the Brownian motion of EVs in suspension, a technique known as nanoparticle-tracking analysis (NTA). This technique is used by many researchers in the EV field. The ability to accurately replicate data between studies and laboratories is critical to advancing the knowledge surrounding EVs for use in liquid biopsy and cancer studies, in general. Thus, this chapter provides a step-by-step guide on isolating EVs using a variety of methods, for characterizing EVs using the NS300 NTA instrument, troubleshooting tips, and a robust guide to reporting key parameters that will help improve cross-lab reproducibility of NTA data.