Ac40ts cantilevers

Glass coverslips were cleaned using a Plasma cleaner Fempto timer with a 40-kHz, 100-W generator (Diener Electronic), and a lipid/sx-1 TMD bilayer was generated by spin-coating as described in Ref. 25 (link). The reconstituted bilayers were imaged with a Cervantes full mode AFM system (Nanotec) using AC40TS cantilevers (f₀ = 110 kHz, k = 0.1 N/m, Olympus) as described previously (28 (link)). Calibration of the cantilevers was accomplished by using the thermal noise spectrum. We employed the Jumping Mode Plus (jump-off, 100 nm; sample points, 50), which allows scanning at controlled vertical forces between 0.2 nN and several nanonewtons (29 (link)).

Imaging and cell elasticity measurements were performed using a Bioscope Resolve atomic force microscope (Bruker, Santa Barbra, CA), operating in the PeakForce live cell mode in DMEM at room temperature. Olympus AC40-TS cantilevers (tip length 40 μm, tip radius 10 nm, nominal spring constant 0.09 N/m) were used in all experiments. Before each measurement, we performed a force-versus-distance curve on the petri dish adjacent to the cells to measure the detection sensitivity and determined the spring constant of the cantilever by recording the thermal noise power spectrum in liquid. For all measurements, the scan rate was 0.3 Hz and 0.4 Hz for whole cell and high resolution imaging of the cortex, respectively, with a frame of 128×128 pixels, using a PeakForce frequency of 1 kHz and an oscillation amplitude of 100 nm. The force setpoint was set to 150 pN and an automatic gain control was used to minimize the PeakForce error. Image analysis was performed using the Nanoscope analysis v1.80 (Bruker Nano Surfaces, Santa Barbara).