KEYWORDS: In vivo imaging, Light sources and illumination, Confocal microscopy, Signal to noise ratio, Optical imaging, Neuroimaging, Crosstalk, Brain, Tissues, Tissue optics
Advancements in genetically encoded voltage indicators (GEVIs) have made it possible to measure cellular membrane potential changes optically. But performing GEVI imaging in vivo remains highly challenging due to factors such as low GEVI concentrations, modest signal dynamic range, tissue scattering, out-of-focus fluorescence. To address these challenges, we developed a microscopy technique that take advatanges from both widefield targeted illumination and confocal background rejection, enabling high SNR low crosstalk GEVI imaging across millimeter fields-of-view, at supra-kilohertz frame rates, over extended durations, and at high penetration depths. We demonstrate our technique under a variety of imaging conditions across multiple brain regions and with different classes of GEVIs.
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