title: All-optical bidirectional neural interfacing using hybrid multiphoton holographic optogenetic stimulation.
abstract: Our understanding of neural information processing could potentially be advanced by combining flexible three-dimensional (3-D) neuroimaging and stimulation. Recent developments in optogenetics suggest that neurophotonic approaches are in principle highly suited for noncontact stimulation of network activity patterns. In particular, two-photon holographic optical neural stimulation (2P-HONS) has emerged as a leading approach for multisite 3-D excitation, and combining it with temporal focusing (TF) further enables axially confined yet spatially extended light patterns. Here, we study key steps toward bidirectional cell-targeted 3-D interfacing by introducing and testing a hybrid new 2P-TF-HONS stimulation path for accurate parallel optogenetic excitation into a recently developed hybrid multiphoton 3-D imaging system. The system is shown to allow targeted all-optical probing of in vitro cortical networks expressing channelrhodopsin-2 using a regeneratively amplified femtosecond laser source tuned to 905 nm. These developments further advance a prospective new tool for studying and achieving distributed control over 3-D neuronal circuits both in vitro and in vivo.
authors.affiliation: Technion-Israel Institute of Technology , Faculty of Biomedical Engineering, Technion City, Haifa 3200000, Israel.
authors.forename: Shir
authors.initials: S
authors.lastname: Paluch-Siegler
authors.affiliation: Technion-Israel Institute of Technology , Faculty of Biomedical Engineering, Technion City, Haifa 3200000, Israel.
authors.forename: Tom
authors.initials: T
authors.lastname: Mayblum
authors.affiliation: Howard Hughes Medical Institute , Janelia Research Campus, Ashburn, Virginia 20147, United States.
authors.forename: Hod
authors.initials: H
authors.lastname: Dana
authors.affiliation: Technion-Israel Institute of Technology , Faculty of Biomedical Engineering, Technion City, Haifa 3200000, Israel.
authors.forename: Inbar
authors.initials: I
authors.lastname: Brosh
authors.affiliation: Ruppin Academic Center , School of Engineering, Medical Engineering, Emeq Hefer 4025000, Israel.
authors.forename: Inna
authors.initials: I
authors.lastname: Gefen
authors.affiliation: Technion-Israel Institute of Technology , Faculty of Biomedical Engineering, Technion City, Haifa 3200000, Israel.
authors.forename: Shy
authors.initials: S
authors.lastname: Shoham
keywords: optogenetics; spatial light modulator; temporal focusing; two-photon microscopy