Single-Shot 3D Widefield Fluorescence Imaging with a Computational Miniature Mesoscope

Abstract

Fluorescence imaging is indispensable to biology and neuroscience. The need for large-scale imaging in freely behaving animals has further driven the development in miniaturized microscopes (miniscopes). However, conventional microscopes / miniscopes are inherently constrained by their limited space-bandwidth-product, shallow depth-of-field, and the inability to resolve 3D distributed emitters. Here, we present a Computational Miniature Mesoscope (CM${}^2$) that overcomes these bottlenecks and enables single-shot 3D imaging across an 8 $\times$ 7-mm${}^2$ field-of-view and 2.5-mm depth-of-field, achieving 7-$\mu$m lateral resolution and better than 200-$\mu$m axial resolution. Notably, the CM${}^2$ has a compact lightweight design that integrates a microlens array for imaging and an LED array for excitation in a single platform. Its expanded imaging capability is enabled by computational imaging that augments the optics by algorithms. We experimentally validate the mesoscopic 3D imaging capability on volumetrically distributed fluorescent beads and fibers. We further quantify the effects of bulk scattering and background fluorescence on phantom experiments.

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