Spatiotemporally decoupled super-oscillatory microscopy enables label-free reconstruction of human ciliary waveforms

Résumé fourni par la source

Imaging native motile cilia requires label-free contrast, sub-diffraction spatial discrimination, and millisecond temporal sampling. We introduce Super-Oscillatory Label-free Inertia-free Scanning (SOLIS) microscopy, in which a static multilevel diffractive optical element generates compressed super-oscillatory probes and a high-speed digital micromirror device addresses the probes without translated-stage motion. A complete 25-position scan produces one acquisition-window-averaged map every 3.58 ms, corresponding to 279 reconstructed fps. On nanofabricated line-pair targets, SOLIS retained measurable label-free contrast at a 253 nm period and resolved representative modulation at 270 nm. In differentiated primary human nasal epithelial cultures, SOLIS distinguished adjacent ciliary maxima separated by 346 nm, below the 431 nm Rayleigh reference, and acquired 14-16 reconstructed samples per observed beat cycle. Tip-centroid tracking recovered closed, asymmetric two-dimensional trajectories with a median cycle-to-cycle localisation precision of 63.5 nm across 20 cilia. In a proof-of-principle perturbation, Dynarrestin reduced culture-level ciliary beat frequency from 19.2 to 6.6 Hz and projected stroke amplitude from 4.8 to 2.3 um. By separating spatial compression from temporal addressing, SOLIS extends super-oscillatory microscopy to label-free, millisecond-scale analysis of human ciliary kinematics.

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