Mfp 3d

Time-lapse imaging was carried out in Asylum MFP3D instrument using the MSNL probe (Bruker Corporation). First the functionalized mica surface was imaged in 10 mM sodium phosphate buffer (pH 7.0) to obtain a clean surface, then the desired amount of protein solution was added (100 nM) and imaging was carried out. On-surface aggregation was monitored by scanning the surface at one hour time intervals. The results shown in Supplementary Fig. 6B have been obtained using Nanoscope Multimode VIII system (Bruker, Santa Barbara, CA) in Peakforce mode. A small droplet of 10 nM alpha-synuclein was created under the fluid cell, then the cell was inserted into scanner head and imaging was being continued after proper alignment of the laser. The imaging was continued in the same area to monitor the changes in morphology of the aggregates.

The TEM images of the BSO-based FTJs were obtained by scanning the specimens using the Titan G2 60–300 instrument, where the specimens were prepared by a focused ion beam tool (FEI Helios NanoLab 600i) operated at the voltage of 2–30 kV. The polarization−electric field (P − E) loops were measured using the ferroelectric measurement system (aixACCT TF-Analyzer 3000) at the frequency of 0.01 − 1 MHz. Atomic force microscopy machines from both Asylum Research (MFP3D) and Bruker (Dimension Icon) were used to obtain the piezoresponse force microscopy images. The box-in-box patterns were written on the BSO film by applying the voltage of +7 V or −7 V to the corresponding regions, respectively. The X-ray diffraction (XRD) pattern of the BSO film was collected by an X-ray diffractometer (PW 3040-X’Pert Pro, PANalytical, Cu Kα, λ = 1.54 Å).

Liquid AFM was performed on an Asylum MFP-3D or Bruker Dimension Icon. In the former, the as provided fluid-cell light was utilised with an 18 mm glass backing to allow for inverted microscopy. The cantilever (DNP-10) was calibrated against a bare glass substrate before sample imaging. The samples were imaged in AC mode, maintaining a maximal setpoint to reduce force during imaging. In the latter, a home-built fluid chamber was constructed atop of a silicon wafer. The cantilever (ScanAsyst-Fluid+) was calibrated against a bare glass substrate in water phase before sample imaging. The samples were imaged in peak force mode with quantitative nanomechanical mapping (PF-QNM). In both cases, the supported lipid bilayers were formed by mimicking the solvent exchange process manually with micropipettes.