A large field‑of‑view,single‑cell‑resolution two‑and three‑photon microscope for deep and wide imaging  

作　　者：Aaron T.Mok Tianyu Wang Shitong Zhao Kristine E.Kolkman Danni Wu Dimitre G.Ouzounov Changwoo Seo Chunyan Wu Joseph R.Fetcho Chris Xu 

机构地区：[1]School of Applied and Engineering Physics,Cornell University,Ithaca,NY,USA [2]Meining School of Biomedical Engineering,Cornell University,Ithaca,NY,USA [3]Department of Electrical and Computer Engineering,Boston University,Boston,MA,USA [4]Department of Neurobiology and Behavior,Cornell University,Ithaca,NY,USA [5]Harvard T.H.Chan School of Public Health,Harvard University,Cambridge,MA,USA [6]Department of Molecular&Cellular Biology,Harvard University,Cambridge,MA,USA [7]McGovern Institute for Brain Research at MIT,Cambridge,MA,USA

出　　处：《eLight》2024年第1期26-39,共14页e光学(英文)

基　　金：National Science Foundation NeuroNex(Grant No.DBI-1707312 to C.X.).NIH/NINDS(Grant No.U01NS103516 to C.X.).Cornell Neurotech Mong Fellowship to A.M.

摘　　要：In vivo imaging of large-scale neuronal activity plays a pivotal role in unraveling the function of the brain’s circuitry.Multiphoton microscopy,a powerful tool for deep-tissue imaging,has received sustained interest in advancing its speed,field of view and imaging depth.However,to avoid thermal damage in scattering biological tissue,field of view decreases exponentially as imaging depth increases.We present a suite of innovations to optimize three-photon microscopy for large field-of-view imaging at depths unreachable by two-photon microscopy.These techniques enable us to image neuronal activities of transgenic animals expressing protein calcium sensors in a~3.5-mm diameter field-of-view with single-cell resolution in the deepest cortical layer of mouse brains.We further demonstrate simultaneous large field-of-view two-photon and three-photon imaging,subcortical imaging in the mouse brain,and whole-brain imaging in adult zebrafish.The demonstrated techniques can be integrated into typical multiphoton microscopes to enlarge field of view for system-level neural circuit research.

关 键 词：Three-photon microscopy Two-photon microscopy Large field of view Brain imaging DEEPscope 

分 类 号：O43[机械工程—光学工程]