Nanoscope software 7

AFM imaging was performed in tapping mode, using the dimension V Bioscope (Bruker, Billerika, MA, USA), equipped with PPP-FMR-SPL silicon tips (Nanosensors, Neuchatel, Switzerland). Briefly, EVs in sucrose buffer (50 µL) were diluted 1:20 in cell and molecular ultrapure sterile water. A fixed amount of properly diluted EV samples (50 μL) was then spin cast onto freshly cleaved mica sheets (grade V-1, thickness of 0.15 mm, size of 15 mm × 15 mm), using a model WS-650SZ-6NPP/LITE spin coater (Laurell, North Wales, PA, USA), at an acceleration of 600 rpm/s, up to a speed of 2000 rpm for 15 s. Imaging was performed using the Nanoscope Software 7.3 (Veeco, Plainview, NY, USA). Scan size ranged from 2.5 to 25 μm, and the scan speed ranged from 5 to 10 μm per second.

AFM imaging was performed using an AFM Dimension V (Bruker, Billerika, MA, USA) equipped with PPP-FMR-SPL silicon tips (Nanosensors, Neuchatel, Switzerland). Briefly, EVs in sucrose buffer (50 µL) and LDL (Sigma-Aldrich, USA and Millipore Corporation, USA) were diluted 1:100 in cell and molecular ultrapure sterile water. A fixed amount of diluted EVs (50 μL) were then deposited onto freshly cleaved mica sheets (grade V-1, thickness of 0.15 mm, size of 15 mm × 15 mm), using a spin cast system model WS-650SZ-6NPP/LITE (Laurell, North Wales, PA, USA) at a speed of 800 relative centrifugal force (rcf) for 15 s. Imaging was performed using the Nanoscope Software 7.3 (Veeco, Plainview, NY, USA). The scan size ranged from 0.5 to 25 μm and the scan speed ranged from 5 μm to 10 μm per second.