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

We here report an aberration corrected 25 plane camera array Multifocus microscope (M25) for high speed, high resolution widefield 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 grating 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.