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Kilohertz frame-rate two-photon tomography

By Abbas Kazemipour, Ondrej Novak, Daniel Flickinger, Jonathan S. Marvin, Jonathan King, Philip Borden, Shaul Druckmann, Karel Svoboda, Loren L. Looger, Kaspar Podgorski

Posted 28 Jun 2018
bioRxiv DOI: 10.1101/357269 (published DOI: 10.1038/s41592-019-0493-9)

Point-scanning two-photon microscopy enables high-resolution imaging within scattering specimens such as the mammalian brain, but sequential acquisition of voxels fundamentally limits imaging speed. We developed a two-photon imaging technique that scans lines of excitation across a focal plane at multiple angles and uses prior information to recover high-resolution images at over 1.4 billion voxels per second. Using a structural image as a prior for recording neural activity, we imaged visually-evoked and spontaneous glutamate release across hundreds of dendritic spines in mice at depths over 250 microns and frame-rates over 1 kHz. Dendritic glutamate transients in anaesthetized mice are synchronized within spatially-contiguous domains spanning tens of microns at frequencies ranging from 1-100 Hz. We demonstrate high-speed recording of acetylcholine and calcium sensors, 3D single-particle tracking, and imaging in densely-labeled cortex. Our method surpasses limits on the speed of raster-scanned imaging imposed by fluorescence lifetime.

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