All computations were performed using the software MATLAB (MathWorks, Natick, MA, USA). The phase-frequency curves recorded at different distances from the sample surface were extracted as ASCII files using Bruker’s Nanoscope Analysis software and loaded into MATLAB. The curves were fitted with a second-order polynomial on the vicinity of π/2 to determine the frequency fπ/2 at different gaps. Then, the frequency shift Δfπ/2 in the bead oscillation was determined by the difference between the acquired frequencies where the phase lag is kept constant at π/2 of the unperturbed sphere placed far from the hair bundle (1 μm) and the perturbed sphere when placed within 50 nm from the top of the tall stereocilia row: Δfπ/2 = fnear,π/2ffar,π/2. In addition, the phase-frequency curves’ slope dϕ/df was computed using the frequency sweeps recorded at 50 nm above the hair bundle by fitting the phase-frequency curves with a third-order polynomial. Estimates of dϕ/df did not change significantly between the measured height range over the sensory cell hair bundle (Fig. 5A).

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