AFM scanning was performed in tapping mode in air with a Titanium atomic force microscope (NT-MDT, Zelenograd, Russia; the microscope pertains to the equipment of “Human Proteome” Core Facility of the Institute of Biomedical Chemistry, supported by Ministry of Education and Science of Russian Federation, agreement 14.621.21.0017, unique project ID: RFMEFI62117X0017) equipped with NSG03 silicon cantilevers with gold reflecting coating (TipsNano, Zelenograd, Russia; typical resonance frequency 47 to 150 kHz, tip curvature radius 10 nm) [37 (link)].
For each scanned sensor area, at least five 5 µm × 5 µm frames with 256 × 256 resolution were obtained. The AFM operation, obtaining AFM images, image processing (flattening correction etc.) and the data export into ASCII format was carried out using standard NOVA Px software (NT-MDT, Zelenograd, Russia) supplied with the atomic force microscope as described elsewhere [33 (link)]. The heights of objects in the AFM images were measured using a specialized software for AFM data processing, developed in the Institute of Biomedical Chemistry (Rospatent registration No. 2010613458). The cut-off level (noise level) amounted to 500 objects per 400 μm2 area [32 (link)]. The height of the AFM-imaged objects was used as the criterion for the determination of their dimensions.
For each scanned sensor area, at least five 5 µm × 5 µm frames with 256 × 256 resolution were obtained. The AFM operation, obtaining AFM images, image processing (flattening correction etc.) and the data export into ASCII format was carried out using standard NOVA Px software (NT-MDT, Zelenograd, Russia) supplied with the atomic force microscope as described elsewhere [33 (link)]. The heights of objects in the AFM images were measured using a specialized software for AFM data processing, developed in the Institute of Biomedical Chemistry (Rospatent registration No. 2010613458). The cut-off level (noise level) amounted to 500 objects per 400 μm2 area [32 (link)]. The height of the AFM-imaged objects was used as the criterion for the determination of their dimensions.
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