High-Speed, Cortex-Wide Volumetric Recording of Neuroactivity at Cellular Resolution using Light Beads Microscopy | bioRxiv

Two-photon microscopy together with genetically encodable calcium indicators has emerged as a standard tool for high-resolution imaging of neuroactivity in scattering brain tissue. However, its various realizations have not overcome the inherent tradeoffs between speed and spatiotemporal sampling in a principled manner which would be necessary to enable, amongst other applications, mesoscale volumetric recording of neuroactivity at cellular resolution and speed compatible with resolving calcium transients. Here, we introduce Light Beads Microscopy (LBM), a scalable and spatiotemporally optimal acquisition approach limited only by fluorescence life-time, where a set of axially-separated and temporally-distinct foci record the entire axial imaging range near-simultaneously, enabling volumetric recording at 1.41 × 108 voxels per second. Using LBM, we demonstrate mesoscopic and volumetric imaging at multiple scales in the mouse cortex, including cellular resolution recordings within ∼3×5×0.5 mm3 volumes containing >200,000 neurons at ∼5 Hz, recording of populations of ∼1 million neurons within ∼5.4×6×0.5 mm3 volumes at ∼2Hz as well as higher-speed (9.6 Hz) sub-cellular resolution volumetric recordings. LBM provides an unprecedented opportunity for discovering the neurocomputations underlying cortex-wide encoding and processing of information in the mammalian brain. ### Competing Interest Statement The authors have declared no competing interest.

High-Speed, Cortex-Wide Volumetric Recording of Neuroactivity at Cellular Resolution using Light Beads Microscopy | bioRxiv Skip to main content New Results High-Speed, Cortex-Wide Volumetric Recording of Neuroactivity at Cellular Resolution using Light Beads Microscopy Jeffrey Demas , Jason Manley , Frank Tejera , Hyewon Kim , Francisca Martínez Traub , Brandon Chen , View ORCID Profile Alipasha Vaziri doi: https://doi.org/10.1101/2021.02.21.432164 Jeffrey Demas 1 Laboratory of Neurotechnology and Biophysics, The Rockefeller University , New York, New York, USA Find this author on Google Scho

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