Pico view1.1 version software

For AFM imaging, purified EVs preparations were diluted with filtered PBS (1:100) and a 5 μl aliquot of the diluted sample solution was deposited on freshly cleaved mica sheet followed by incubation at room temperature for 10 min. The dried sample was then gently washed with 200 μl of Milli-Q water to remove salt and loosely bound moieties. AFM experiments were performed in intermittent contact mode (called “tapping” or AAC mode) to minimize tip-induced damage. AAC mode AFM was performed using a Pico plus 5500 inverted light microscope AFM (Agilent Technologies) with a Piezo scanner (maximum range 9 μm). Microfabricated silicon cantilevers 225 μm in length with a nominal spring force constant of 21 to 98 N/m were used (Nano Sensors). Cantilever oscillation frequency was tuned into resonance frequency. The cantilever resonance frequency was 150 to 300 kHz. The images (512 × 512 pixels) were captured with a scan size between 0.5 and 0.8 μm at a scan speed rate of 0.5 lines/s. Images were processed by flatten using Pico view1.1 version software (Agilent Technologies). Image manipulation was done using Pico Image Advanced version software (Agilent Technologies).