In vivo voltage imaging with targeted illumination confocal (TICO) microscopy

Sheng Xiao; William J. Cunningham; Krishnakanth Kondabolu; Eric Lowet; Maria V. Moya; Rebecca Mount; Cara Ravasio; Michael N. Economo; Xue Han; Jerome Mertz

doi:10.1117/12.3000992

13 March 2024 In vivo voltage imaging with targeted illumination confocal (TICO) microscopy

Sheng Xiao, William J. Cunningham, Krishnakanth Kondabolu, Eric Lowet, Maria V. Moya, Rebecca Mount, Cara Ravasio, Michael N. Economo, Xue Han, Jerome Mertz

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Proceedings Volume PC12828, Neural Imaging and Sensing 2024; PC1282804 (2024) https://doi.org/10.1117/12.3000992
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

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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Sheng Xiao, William J. Cunningham, Krishnakanth Kondabolu, Eric Lowet, Maria V. Moya, Rebecca Mount, Cara Ravasio, Michael N. Economo, Xue Han, and Jerome Mertz "In vivo voltage imaging with targeted illumination confocal (TICO) microscopy", Proc. SPIE PC12828, Neural Imaging and Sensing 2024, PC1282804 (13 March 2024); https://doi.org/10.1117/12.3000992

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KEYWORDS

In vivo imaging

Confocal microscopy

Light sources and illumination

Signal to noise ratio

Brain

Crosstalk

Neuroimaging

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