High-speed 3D Imaging with 25-Camera Multifocus Microscope

Hirata-Miyasaki, E.; Bajor, A. A.; Petterson, G. M.; Senftleben, M. L.; Fouke, K. E.; Graham, T. G. W.; John, D. D.; Morgan, J. R.; Haspel, G.; Abrahamsson, S.

doi:10.1101/2024.09.23.614351

bioRxiv · 10.1101/2024.09.23.614351

High-speed 3D Imaging with 25-Camera Multifocus Microscope

Hirata-Miyasaki, E.·Bajor, A. A.·Petterson, G. M.·Senftleben, M. L.·Fouke, K. E.·Graham, T. G. W.·John, D. D.·Morgan, J. R.·Haspel, G.·Abrahamsson, S.

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Abstract

We here report an aberration-corrected 25-plane camera array Multifocus microscope (M25) for high-speed, high-resolution wide-field optical microscopy in three spatial dimensions (3D). We demonstrate live imaging of 25-plane 3D volumes of up to 180x180x50um at >100 volumes per second. 3D data is recorded simultaneously by an array of 25 small, sensitive, synchronized machine-vision cameras. M25 employs aberration-corrected Multifocus microscopy--an optical method where diffractive Fourier optics are used for multiplexing and refocusing light-- with a simplified design for chromatic dispersion correction where a corrective diffractive gratings is placed on each camera in the array. This elegant architecture for chromatic correction will be applicable in a broad range of diffractive imaging applications. M25 is a powerful optical tool for high-speed 3D microscopy in that it allows both non-invasive, label-free bright-field and highly sensitive fluorescence microscopy. We showcase M25 capabilities in 3D particle tracking, bright-field, and fluorescence imaging in D. melanogaster, and locomotion and neural activity studies in C. elegans.

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Hirata-Miyasaki, E., Bajor, A. A., Petterson, G. M., Senftleben, M. L., Fouke, K. E., Graham, T. G. W., John, D. D., Morgan, J. R., Haspel, G., Abrahamsson, S.. 2024-09-24. High-speed 3D Imaging with 25-Camera Multifocus Microscope. https://doi.org/10.1101/2024.09.23.614351

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