Np100

TRPS measurements were performed with the qNano (Izon Science, UK). The instrument was set up and calibrated as per manufacturer recommendations. For liposome samples, a polyurethane nanopore rated for particles between 100 and 250 nm (NP150, Izon Science, UK) was used to perform all measurements, and was axially stretched to 48 mm, as measured from adjacent teeth on the qNano unit. For exosome samples, a polyurethane nanopore rated for particles <100 nm (NP100-, Izon Science, UK) was used, and was axially stretched to 48 mm, as measured from adjacent teeth on the qNano unit. 40 µL of sample diluted to an appropriate particle content in PBS + 0.05% Tween-20 was measured with this system. Optimally, measurement durations were greater than two minutes except where system instability limited this. All measurements were calibrated with 115 nm (NP100-) or 212 nm (NP150) polystyrene beads appropriately diluted (Izon Science, UK). Data processing and analysis were carried out on the Izon Control Suite software v2.2 (Izon Science, UK). Pore sensitivity was defined by the smallest possible particle able to be seen above system noise (i.e. pulse magnitudes >0.05 nA), as detailed in the following work (paper in review).

EVs were resuspended in PBS, and the size and number of particles were analysed using qNano (Izon, Christchurch, New Zealand). The NP100 nanopore (Izon) was used for detecting particles, and the measuring system was calibrated using CPC100 calibration particles (Izon). Particles were measured at 47.0 mm stretch at 0.7 V.