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Seeing in the Dark: Intelligent Fourier Light Field Imaging for Bioluminescence Microscopy

Krieg, M.; Morales-Curiel, L.-F.

2025-10-14 biophysics Community evaluation
10.1101/2025.10.13.680966 bioRxiv
Show abstract

Bioluminescence microscopy offers a uniquely non-invasive window into cellular dynamics, yet its use has traditionally been limited by the intrinsically low brightness of luciferases. The poor photon budget forces long exposures, preventing faithful visualization of rapid physiological processes, especially in three dimensions. To overcome this barrier, we developed a Fourier light field microscope coupled with deep-learning-based reconstruction that achieves sub-second volumetric bioluminescence imaging with significantly improved spatial resolution. This approach eliminates the speed-resolution trade-off of conventional light field methods and bypasses the need for slow classical deconvolution. We demonstrate its power by performing real-time 3D calcium imaging in freely moving Caenorhabditis elegans, and by quantifying cell dynamics within stem-cell-derived spheroids using fluorescently labeled nuclei and calcium dynamics in muscles and neurons. Together, these results establish our framework as a practical tool for dynamic, volumetric studies of living systems.

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