Optics-aware super-resolution light-field microscopy for long-term volumetric imaging of dynamic intracellular processes at millisecond timescales

Zhu, L.; Sun, J.; Yi, C.; Zhang, M.; Niu, G.; Tang, J.; Zhang, Y.; Li, D.; Fei, P.

doi:10.1101/2023.03.15.532876

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bioRxiv · 10.1101/2023.03.15.532876

Optics-aware super-resolution light-field microscopy for long-term volumetric imaging of dynamic intracellular processes at millisecond timescales

Zhu, L.·Sun, J.·Yi, C.·Zhang, M.·Niu, G.·Tang, J.·Zhang, Y.·Li, D.·Fei, P.

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Abstract

Long-term and high-spatiotemporal-resolution 3D imaging of living cells remains an unmet challenge for super-resolution microscopy, owing to the noticeable phototoxicity and limited scanning speed. While emerging light-field microscopy can mitigate this issue through three-dimensionally capturing biological dynamics with merely single snapshot, it suffers from suboptimal resolution insufficient for resolving subcellular structures. Here we propose an Adaptive Learning PHysics-Aware Light-Field Microscopy (Alpha-LFM) with a physics-aware deep learning framework and adaptive-tuning strategies capable for highly-generalizable light-field reconstruction of diverse subcellular dynamics. Alpha-LFM delivers sub-diffraction-limit spatial resolution ([~]120 nm) while maintaining high temporal resolution and low phototoxicity. It enables rapid (at hundreds of volumes per second), long-term (up to 60 hours) 3D super-resolution imaging of diverse intracellular dynamics with exceptional details. Using Alpha-LFM approach, we finely resolve the lysosome-mitochondrial interactions, capture rapid motion of peroxisome and the endoplasmic reticulum, and reveal the variations in mitochondrial fission activity throughout two complete cell cycles.

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Zhu, L., Sun, J., Yi, C., Zhang, M., Niu, G., Tang, J., Zhang, Y., Li, D., Fei, P.. 2023-03-16. Optics-aware super-resolution light-field microscopy for long-term volumetric imaging of dynamic intracellular processes at millisecond timescales. https://doi.org/10.1101/2023.03.15.532876

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