Nanosight lm10 nta instrument

The particle size and concentration were measured by nanoparticle tracking analysis (NTA) using a NanoSight LM10 NTA instrument with NTA 3.2 software for data acquisition and analysis (Malvern Panalytical). The pEXOs samples were diluted with filtered PBS to match the recommended particle concentration range {20–120 particles/field; [(1 × 10⁷)–(1 × 10⁹)] particles/ml}. Three 60-s videos were acquired for each sample maintaining a camera level, 10; and the videos were analyzed at detection threshold, 3. The modal particle size and particles concentration/ml was measured by the software. The pEXOs concentration was inferred from the particle size distribution up to 150-nm size, it was normalized by resuspension volume and expressed relative to the starting plasma volume.

Nanoparticle tracking analysis (NTA) was performed to validate EV shape, as well as elucidate their concentration, and provide an orthogonal measure of EV size. Briefly, isolated EVs were resuspended in sterile filtered PBS and passed through a 0.2 μm filter. Analysis was then performed on a Nanosight LM10 NTA instrument (Malvern Panalytical) calibrated to 100 nm latex calibration beads. Videos were generated from all biological replicates in this experiment, as well as size characterization and particulate concentration measurements.