High-Speed Low-Light In Vivo Two-Photon Voltage Imaging of Large Neuronal Populations
High-Speed Low-Light In Vivo Two-Photon Voltage Imaging of Large Neuronal Populations
SUMMARY Monitoring spiking activity across large neuronal populations at behaviorally relevant timescales is critical for understanding neural circuit function. Unlike calcium imaging, voltage imaging requires kilohertz sampling rates which reduces fluorescence detection to near shot noise levels. High-photon flux excitation can overcome photon-limited shot noise but photo-bleaching and photo-damage restricts the number and duration of simultaneously imaged neurons. We investigated an alternative approach aimed at low two-photon flux, voltage imaging below the shot noise limit. This framework involved developing: a positive-going voltage indicator with improved spike detection (SpikeyGi); an ultra-fast two-photon microscope for kilohertz frame-rate imaging across a 0.4×0.4mm2 field of view, and; a self-supervised denoising algorithm (DeepVID) for inferring fluorescence from shot-noise limited signals. Through these combined advances, we achieved simultaneous high-speed, deep-tissue imaging of more than one hundred densely-labeled neurons over one hour in awake behaving mice. This demonstrates a scalable approach for voltage imaging across increasing neuronal populations.
Ahrens Allison M.、Tian Lei、Platisa Jelena、Ye Xin、Chen Ichun Anderson、Davison Ian G.、Pieribone Vincent A.、Chen Jerry L.、Liu Chang
Department of Biology, Boston UniversityDepartment of Biomedical Engineering, Boston University||Center for Neurophotonics, Boston University||Department of Electrical and Computer Engineering, Boston UniversityDepartment of Cellular and Molecular Physiology, Yale University||Department of Neuroscience, Yale University||The John B. Pierce LaboratoryDepartment of Biomedical Engineering, Boston University||Center for Neurophotonics, Boston UniversityCenter for Neurophotonics, Boston UniversityCenter for Neurophotonics, Boston University||Department of Biology, Boston University||Center for Systems Neuroscience, Boston UniversityDepartment of Cellular and Molecular Physiology, Yale University||Department of Neuroscience, Yale University||The John B. Pierce LaboratoryDepartment of Biomedical Engineering, Boston University||Center for Neurophotonics, Boston University||Department of Biology, Boston University||Center for Systems Neuroscience, Boston UniversityDepartment of Electrical and Computer Engineering, Boston University
生物科学研究方法、生物科学研究技术生物物理学光电子技术
Ahrens Allison M.,Tian Lei,Platisa Jelena,Ye Xin,Chen Ichun Anderson,Davison Ian G.,Pieribone Vincent A.,Chen Jerry L.,Liu Chang.High-Speed Low-Light In Vivo Two-Photon Voltage Imaging of Large Neuronal Populations[EB/OL].(2025-03-28)[2025-06-07].https://www.biorxiv.org/content/10.1101/2021.12.07.471668.点此复制
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