Guorong Hu

Wide-field fluorescence microscopy with compact optics often suffers from spatially varying blur due to field-dependent aberrations, vignetting, and sensor truncation, while finite sensor sampling imposes an inherent trade-off between field of view (FOV) and resolution. Computational Miniaturized Mesoscope (CM2) alleviate the sampling limit by multiplexing multiple sub-views onto a single sensor, but introduce view crosstalk and a highly ill-conditioned inverse problem compounded by spatially variant point spread functions (PSFs). Prior learning-based spatially varying (SV) reconstruction methods typically rely on global SV operators with fixed input sizes, resulting in memory and training costs that scale poorly with image dimensions. We propose SV-CoDe (Spatially Varying Coordinate-conditioned Deconvolution), a scalable deep learning framework that achieves uniform, high-resolution reconstruction across a 6.5 mm FOV. Unlike conventional methods, SV-CoDe employs coordinate-conditioned convolutions to locally adapt reconstruction kernels; this enables patch-based training that decouples parameter count from FOV size. SV-CoDe achieves the best image quality in both simulated and experimental measurements while requiring 10x less model size and 10x less training data than prior baselines. Trained purely on physics-based simulations, the network robustly generalizes to bead phantoms, weakly scattering brain slices, and freely moving C. elegans. SV-CoDe offers a scalable, physics-aware solution for correcting SV blur in compact optical systems and is readily extendable to a broad range of biomedical imaging applications.

Ultrafast 3D imaging is indispensable for visualizing complex and dynamic biological processes. Conventional scanning-based techniques necessitate an inherent trade-off between acquisition speed and space-bandwidth product (SBP). Emerging single-shot 3D wide-field techniques offer a promising alternative but are bottlenecked by the synchronous readout constraints of conventional CMOS systems, thus restricting data throughput to maintain high SBP at limited frame rates. To address this, we introduce EventLFM, a straightforward and cost-effective system that overcomes these challenges by integrating an event camera with Fourier light field microscopy (LFM), a state-of-the-art single-shot 3D wide-field imaging technique. The event camera operates on a novel asynchronous readout architecture, thereby bypassing the frame rate limitations inherent to conventional CMOS systems. We further develop a simple and robust event-driven LFM reconstruction algorithm that can reliably reconstruct 3D dynamics from the unique spatiotemporal measurements captured by EventLFM. Experimental results demonstrate that EventLFM can robustly reconstruct fast-moving and rapidly blinking 3D fluorescent samples at kHz frame rates. Furthermore, we highlight EventLFM's capability for imaging of blinking neuronal signals in scattering mouse brain tissues and 3D tracking of GFP-labeled neurons in freely moving C. elegans. We believe that the combined ultrafast speed and large 3D SBP offered by EventLFM may open up new possibilities across many biomedical applications.

SignificanceFluorescence head-mounted microscopes, i.e., miniscopes, have emerged as powerful tools to analyze in-vivo neural populations but exhibit a limited depth-of-field (DoF) due to the use of high numerical aperture (NA) gradient refractive index (GRIN) objective lenses.AimWe present extended depth-of-field (EDoF) miniscope, which integrates an optimized thin and lightweight binary diffractive optical element (DOE) onto the GRIN lens of a miniscope to extend the DoF by 2.8 × between twin foci in fixed scattering samples.ApproachWe use a genetic algorithm that considers the GRIN lens’ aberration and intensity loss from scattering in a Fourier optics-forward model to optimize a DOE and manufacture the DOE through single-step photolithography. We integrate the DOE into EDoF-Miniscope with a lateral accuracy of 70 μm to produce high-contrast signals without compromising the speed, spatial resolution, size, or weight.ResultsWe characterize the performance of EDoF-Miniscope across 5- and 10-μm fluorescent beads embedded in scattering phantoms and demonstrate that EDoF-Miniscope facilitates deeper interrogations of neuronal populations in a 100-μm-thick mouse brain sample and vessels in a whole mouse brain sample.ConclusionsBuilt from off-the-shelf components and augmented by a customizable DOE, we expect that this low-cost EDoF-Miniscope may find utility in a wide range of neural recording applications.