All-Optical Volumetric Physiology for Connectomics in Dense Neuronal Structures

Summary All-optical physiology (AOP) manipulates and reports neuronal activities with light, allowing for interrogation of neuronal functional connections with high spatiotemporal resolution. However, contemporary high-speed AOP platforms are limited to single-depth or discrete multi-plane recordings that are not suitable for studying functional connections among densely packed small neurons, such as neurons in Drosophila brains. Here, we constructed a 3D AOP platform by incorporating single-photon point stimulation and two-photon high-speed volumetric recordings with a tunable acoustic gradient-index (TAG) lens. We demonstrated the platform effectiveness by studying the anterior visual pathway (AVP) of Drosophila. We achieved functional observation of spatiotemporal coding and the strengths of calcium-sensitive connections between anterior optic tubercle (AOTU) sub-compartments and >70 tightly assembled 2-μm bulb (BU) microglomeruli in 3D coordinates with a single trial. Our work aids the establishment of in vivo 3D functional connectomes in neuron-dense brain areas.

[1]  K. Svoboda,et al.  Channelrhodopsin-2–assisted circuit mapping of long-range callosal projections , 2007, Nature Neuroscience.

[2]  Adam S. Charles,et al.  Volumetric Two-photon Imaging of Neurons Using Stereoscopy (vTwINS) , 2016, Nature Methods.

[3]  P. Demoly,et al.  [Transgenic mice]. , 1992, Annales de dermatologie et de venereologie.

[4]  Pál Maák,et al.  Fast 3D Imaging of Spine, Dendritic, and Neuronal Assemblies in Behaving Animals. , 2016, Neuron.

[5]  F. Helmchen,et al.  Imaging cellular network dynamics in three dimensions using fast 3D laser scanning , 2007, Nature Methods.

[6]  M. Häusser,et al.  All-Optical Interrogation of Neural Circuits , 2015, The Journal of Neuroscience.

[7]  Johannes D. Seelig,et al.  Video-rate volumetric functional imaging of the brain at synaptic resolution , 2016, Nature Neuroscience.

[8]  Thomas Hummel,et al.  A topographic visual pathway into the central brain of Drosophila , 2017 .

[9]  Keith J. Kelleher,et al.  Three-dimensional random access multiphoton microscopy for functional imaging of neuronal activity , 2008, Nature Neuroscience.

[10]  M. Häusser,et al.  Targeting neurons and photons for optogenetics , 2013, Nature Neuroscience.

[11]  Shawn R. Olsen,et al.  Cracking neural circuits in a tiny brain: new approaches for understanding the neural circuitry of Drosophila , 2008, Trends in Neurosciences.

[12]  Eirini Papagiakoumou,et al.  Optical developments for optogenetics , 2013, Biology of the cell.

[13]  Ann-Shyn Chiang,et al.  Parallel Neural Pathways Mediate CO2 Avoidance Responses in Drosophila , 2013, Science.

[14]  Weijian Yang,et al.  In vivo imaging of neural activity , 2017, Nature Methods.

[15]  Michael Häusser,et al.  Simultaneous all-optical manipulation and recording of neural circuit activity with cellular resolution in vivo , 2014, Nature Methods.

[16]  W. Denk,et al.  Two-photon laser scanning fluorescence microscopy. , 1990, Science.

[17]  E. Papagiakoumou,et al.  Two-photon optogenetics. , 2012, Progress in brain research.

[18]  D. Tank,et al.  Two-photon excitation of channelrhodopsin-2 at saturation , 2009, Proceedings of the National Academy of Sciences.

[19]  W. Denk,et al.  Deep tissue two-photon microscopy , 2005, Nature Methods.

[20]  David S. Koos,et al.  Deep and fast live imaging with two-photon scanned light-sheet microscopy , 2011, Nature Methods.

[21]  Shun-Chi Wu,et al.  Millisecond two-photon optical ribbon imaging for small-animal functional connectome study. , 2019, Optics letters.

[22]  J. Tiago Gonçalves,et al.  Simultaneous 2-photon calcium imaging at different cortical depths in vivo with spatiotemporal multiplexing , 2010, Nature Methods.

[23]  W. C. Hall,et al.  High-speed mapping of synaptic connectivity using photostimulation in Channelrhodopsin-2 transgenic mice , 2007, Proceedings of the National Academy of Sciences.

[24]  O. Paulsen,et al.  Aberration-free three-dimensional multiphoton imaging of neuronal activity at kHz rates , 2012, Proceedings of the National Academy of Sciences.

[25]  J. Lichtman,et al.  Optical sectioning microscopy , 2005, Nature Methods.

[26]  Liangyi Chen,et al.  Large-field high-resolution two-photon digital scanned light-sheet microscopy , 2014, Cell Research.

[27]  Jerome Mertz,et al.  Wide-field fluorescence sectioning with hybrid speckle and uniform-illumination microscopy. , 2008, Optics letters.

[28]  R. Prevedel,et al.  Brain-wide 3D imaging of neuronal activity in Caenorhabditis elegans with sculpted light , 2013, Nature Methods.

[29]  Johannes D. Seelig,et al.  Feature detection and orientation tuning in the Drosophila central complex , 2013, Nature.

[30]  M. Häusser,et al.  Targeted single-cell electroporation of mammalian neurons in vivo , 2009, Nature Protocols.

[31]  Louis K. Scheffer,et al.  A visual motion detection circuit suggested by Drosophila connectomics , 2013, Nature.

[32]  Volker Hartenstein,et al.  Visual Input to the Drosophila Central Complex by Developmentally and Functionally Distinct Neuronal Populations , 2017, Current Biology.

[33]  Stefanie Hampel,et al.  Targeted Manipulation of Neuronal Activity in Behaving Adult Flies , 2017 .

[34]  R. Yuste,et al.  The Brain Activity Map Project and the Challenge of Functional Connectomics , 2012, Neuron.

[35]  Benjamin F. Grewe,et al.  Two-photon optogenetic toolbox for fast inhibition, excitation and bistable modulation , 2012, Nature Methods.

[36]  Martí Duocastella,et al.  Simultaneous imaging of multiple focal planes for three-dimensional microscopy using ultra-high-speed adaptive optics. , 2012, Journal of biomedical optics.

[37]  Willy Supatto,et al.  Whole-brain functional imaging with two-photon light-sheet microscopy , 2015, Nature Methods.

[38]  Guan-Yu Chen,et al.  Three-Dimensional Reconstruction of Brain-wide Wiring Networks in Drosophila at Single-Cell Resolution , 2011, Current Biology.

[39]  A. Bègue,et al.  Three-dimensional imaging and photostimulation by remote-focusing and holographic light patterning , 2011, Proceedings of the National Academy of Sciences.

[40]  Jeremy Freeman,et al.  Technologies for imaging neural activity in large volumes , 2016, Nature Neuroscience.

[41]  Jeffrey N. Stirman,et al.  Wide field-of-view, multi-region two-photon imaging of neuronal activity in the mammalian brain , 2016, Nature Biotechnology.

[42]  T. Murphy,et al.  Automated light-based mapping of motor cortex by photoactivation of channelrhodopsin-2 transgenic mice , 2009, Nature Methods.

[43]  Kenji F. Tanaka,et al.  Near-infrared deep brain stimulation via upconversion nanoparticle–mediated optogenetics , 2018, Science.

[44]  Yanmeng Guo,et al.  Precise Spatiotemporal Control of Optogenetic Activation Using an Acousto-Optic Device , 2011, PloS one.

[45]  W. Webb,et al.  Nonlinear magic: multiphoton microscopy in the biosciences , 2003, Nature Biotechnology.

[46]  J. Sanes,et al.  Ome sweet ome: what can the genome tell us about the connectome? , 2008, Current Opinion in Neurobiology.

[47]  John G. Flanagan,et al.  Development of Continuous and Discrete Neural Maps , 2007, Neuron.

[48]  Stefan R. Pulver,et al.  Genetically encoded calcium indicators for multi-color neural activity imaging and combination with optogenetics , 2013, Front. Mol. Neurosci..

[49]  Laura Waller,et al.  Precise multimodal optical control of neural ensemble activity , 2018, Nature Neuroscience.

[50]  Valentina Emiliani,et al.  Fast Calcium Imaging with Optical Sectioning via HiLo Microscopy , 2015, PloS one.

[51]  Charles P. Lin,et al.  Continuous volumetric imaging via an optical phase-locked ultrasound lens , 2015, Nature Methods.

[52]  Benjamin F. Grewe,et al.  Fast two-layer two-photon imaging of neuronal cell populations using an electrically tunable lens , 2011, Biomedical optics express.

[53]  Herwig Baier,et al.  Linking Neurons to Network Function and Behavior by Two-Photon Holographic Optogenetics and Volumetric Imaging , 2017, Neuron.

[54]  Jerome Mertz,et al.  Optically sectioned in vivo imaging with speckle illumination HiLo microscopy. , 2011, Journal of biomedical optics.

[55]  Alberto Diaspro,et al.  Enhanced volumetric imaging in 2‐photon microscopy via acoustic lens beam shaping , 2018, Journal of biophotonics.

[56]  Alberto Diaspro,et al.  Simultaneous multiplane confocal microscopy using acoustic tunable lenses. , 2014, Optics express.

[57]  Ling Fu,et al.  All-optical imaging and manipulation of whole-brain neuronal activities in behaving larval zebrafish. , 2018, Biomedical optics express.

[58]  David W. Tank,et al.  Regression-Based Identification of Behavior-Encoding Neurons During Large-Scale Optical Imaging of Neural Activity at Cellular Resolution , 2010, Journal of neurophysiology.

[59]  Ann-Shyn Chiang,et al.  Optical properties of adult Drosophila brains in one-, two-, and three-photon microscopy. , 2019, Biomedical optics express.

[60]  P. Bonifazi,et al.  Simultaneous high-speed imaging and optogenetic inhibition in the intact mouse brain , 2017, Scientific Reports.

[61]  Philipp J. Keller,et al.  Whole-brain functional imaging at cellular resolution using light-sheet microscopy , 2013, Nature Methods.

[62]  Karel Svoboda,et al.  From cudgel to scalpel: toward precise neural control with optogenetics , 2011, Nature Methods.

[63]  E. Isacoff,et al.  Scanless two-photon excitation of channelrhodopsin-2 , 2010, Nature Methods.

[64]  R. Yuste,et al.  Simultaneous two-photon imaging and two-photon optogenetics of cortical circuits in three dimensions , 2018, eLife.

[65]  Laura Waller,et al.  Three-dimensional scanless holographic optogenetics with temporal focusing (3D-SHOT) , 2017, Nature Communications.

[66]  E. Marder,et al.  From the connectome to brain function , 2013, Nature Methods.

[67]  Jerome Mertz,et al.  Optical sectioning microscopy with planar or structured illumination , 2011, Nature Methods.

[68]  Alexandre Mermillod-Blondin,et al.  Two-photon microscopy with simultaneous standard and extended depth of field using a tunable acoustic gradient-index lens. , 2009, Optics letters.

[69]  Valentina Emiliani,et al.  Recent advances in patterned photostimulation for optogenetics , 2017 .

[70]  M. Häusser,et al.  Electrophysiology in the age of light , 2009, Nature.

[71]  Nathan R. Wilson,et al.  Division and subtraction by distinct cortical inhibitory networks in vivo , 2012, Nature.

[72]  Hokto Kazama,et al.  Parallel encoding of recent visual experience and self-motion during navigation in Drosophila , 2017, Nature Neuroscience.

[73]  Balázs Rózsa,et al.  Fast two-photon in vivo imaging with three-dimensional random-access scanning in large tissue volumes , 2012, Nature Methods.

[74]  D. Tank,et al.  Simultaneous cellular-resolution optical perturbation and imaging of place cell firing fields , 2014, Nature Neuroscience.

[75]  Valentina Emiliani,et al.  Three-dimensional spatiotemporal focusing of holographic patterns , 2016, Nature Communications.

[76]  Stefan R. Pulver,et al.  Ultra-sensitive fluorescent proteins for imaging neuronal activity , 2013, Nature.

[77]  Stefan R. Pulver,et al.  Genetically encoded calcium indicators for multi-color neural activity imaging and combination with optogenetics , 2013, Front. Mol. Neurosci..

[78]  G. Ellis‐Davies,et al.  In vivo two‐photon uncaging of glutamate revealing the structure–function relationships of dendritic spines in the neocortex of adult mice , 2011, The Journal of physiology.

[79]  Hitoshi Sakano,et al.  Topographic mapping--the olfactory system. , 2010, Cold Spring Harbor perspectives in biology.

[80]  Aljoscha Nern,et al.  Neural signatures of dynamic stimulus selection in Drosophila , 2017, Nature Neuroscience.

[81]  L. Paninski,et al.  Simultaneous Multi-plane Imaging of Neural Circuits , 2016, Neuron.

[82]  Alberto Diaspro,et al.  Fast Inertia-Free Volumetric Light-Sheet Microscope , 2017 .

[83]  Kevin M. Dean,et al.  Uniform and scalable light-sheets generated by extended focusing. , 2014, Optics express.

[84]  Samer Hattar,et al.  Architecture of retinal projections to the central circadian pacemaker , 2016, Proceedings of the National Academy of Sciences.

[85]  Ann-Shyn Chiang,et al.  Optimizing depth-of-field extension in optical sectioning microscopy techniques using a fast focus-tunable lens. , 2017, Optics express.

[86]  George J. Augustine,et al.  Optogenetic probing of functional brain circuitry , 2011, Experimental physiology.