Qnano software

For determination of the long-term stability of SAMiRNA, the nanoparticle size was monitored by qNano Gold (Izon Science) according to standard operating procedures. Briefly, 35 μl of SAMiRNA-AR68 was analyzed with qNano Gold equipment using an NP80 Nanopore (Izon Science) and applying a 47 mm stretch, a current of 140 nA, and 10 mBar parametric conditions. The calibration particles (CPC100, Izon Science) were assayed before the experimental samples under identical conditions. Particle counts (≥ 50 events each) were finally determined using the qNano software provided by Izon Science (Izon Control Suite Version 3.3).

NTA was performed using the TRPS technique and analyzed on a qNano Gold system (Izon, Medford, MA) to determine the size and concentration of EV particles. With the qNano instrument, an electric current between the two fluid chambers is disrupted when a particle passes through a nanopore NP150 with an analysis range of 70 nm to 420 nm, causing a blockade event to be recorded. The magnitude of the event is proportional to the amount of particles traversing the pore, and the blockade rate directly relates to particle concentration that is measured particle by particle. The results can be calibrated using a single-point calibration under the same measurement conditions used for EV particles (stretch, voltage, and pressure) (80 (link)). CPC200 calibration particles (Izon) were diluted in filtered Izon-supplied electrolyte at 1:500 to equilibrate the system prior to measuring EVs and for useused as a reference. Isolated extracellular vesicle samples were diluted at 0, 1:10, 1:100, and 1:1,000. For the measurements, a 35 µl sample was added to the upper fluid chamber and two working pressures, 5 mbar and 10 mbar, were applied under a current of 120 nA. Particle rates between 200 and 1500 particles/min were obtained. The size, density, and distribution of the particles were analyzed using qNano software (Izon).