Fig. 4: Dynamic analysis of the rotary mechanism via single-particle TIRFM.

a Sketch of the experimental set-up: the mechanism is fixed to the cover slide via biotin–neutravidin–biotin bridges. The crossbar of the camshaft is lengthened to approximately 290 nm. Inset: top view. b Typical field of view image (left, scale bar: 5 µm) and images of the standard deviation over all frames of single-particle movies for exemplary particles (right, scale bar: 600 nm). c Exemplary sequence of frames of a typical particle jumping between three preferred positions (blue, orange, and yellow). d Exemplary angle-time trace of a single particle. e Cumulative angle over time for two typical particles. f Probability density distribution for lever orientations computed from angle-time trace of a typical particle. g Solid line: Distribution of the measured distance of lever arm tip from center averaged over N = 212 particles. Dashed lines: plus/minus standard deviation. Vertical gray line: weighted average of the distance from center. h Solid line: angular velocity distribution computed from N = 212 particles. Dashed lines: plus/minus standard deviation. Vertical gray line indicates the weighted average of the angular velocity (which is zero, since there is no directional bias). i Solid line: average angular root mean square deviation over time from N = 212 particles. Dashed lines: plus/minus standard deviation. Red dashed line: fit using \({{{{{\rm{RMSD}}}}}}={A}\sqrt{{t}}\). Source data are provided as a Source data file.