Two-Color Volumetric Imaging of Neuronal Activity of Cortical Columns

SUMMARY To capture the emergent properties of neural circuits, high-speed volumetric imaging of neural activity at cellular resolution is needed. Here, we introduce wavelength multiplexing to perform fast volumetric two-photon imaging of cortical columns (>2,000 neurons in 10 planes at 10 vol/s), using two different calcium indicators, an electrically tunable lens and a spatial light modulator. We image the activity of neuronal populations from layers 2/3 to 5 of primary visual cortex from awake mice, finding a lack of columnar structures in orientation responses and revealing correlations between layers which differ from trial to trial. We also simultaneously image functional correlations between presynaptic layer 1 axons and postsynaptic layer 2/3 neurons. Wavelength multiplexing enhances high-speed volumetric microscopy and can be combined with other optical multiplexing methods to easily boost imaging throughput.

[1]  Christopher J. Cueva,et al.  Natural Grouping of Neural Responses Reveals Spatially Segregated Clusters in Prearcuate Cortex , 2015, Neuron.

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

[3]  K. Svoboda,et al.  A Cellular Resolution Map of Barrel Cortex Activity during Tactile Behavior , 2015, Neuron.

[4]  Y. Dan,et al.  Clonally Related Visual Cortical Neurons Show Similar Stimulus Feature Selectivity , 2012, Nature.

[5]  Rafael Yuste,et al.  moco: Fast Motion Correction for Calcium Imaging , 2015, Front. Neuroinform..

[6]  Santo Fortunato,et al.  Consensus clustering in complex networks , 2012, Scientific Reports.

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

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

[9]  G. Love,et al.  Wave-front correction and production of Zernike modes with a liquid-crystal spatial light modulator. , 1997, Applied Optics.

[10]  C. Gilbert,et al.  Brain States: Top-Down Influences in Sensory Processing , 2007, Neuron.

[11]  David Pfau,et al.  Simultaneous Denoising, Deconvolution, and Demixing of Calcium Imaging Data , 2016, Neuron.

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

[13]  K. Fuxe,et al.  Hypothalamic Vasopressinergic Projections Innervate Central Amygdala GABAergic Neurons: Implications for Anxiety and Stress Coping , 2016, Front. Neural Circuits.

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

[15]  Delbert Dueck,et al.  Clustering by Passing Messages Between Data Points , 2007, Science.

[16]  R. Yuste,et al.  Visual stimuli recruit intrinsically generated cortical ensembles , 2014, Proceedings of the National Academy of Sciences.

[17]  M. Carandini.  Amplification of Trial-to-Trial Response Variability by Neurons in Visual Cortex , 2004, PLoS biology.

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

[19]  Daniel N. Hill,et al.  Development of Direction Selectivity in Mouse Cortical Neurons , 2011, Neuron.

[20]  Ying Ma,et al.  Penalized matrix decomposition for denoising, compression, and improved demixing of functional imaging data , 2018, bioRxiv.

[21]  G. Gerstein,et al.  Trial-to-Trial Variability and State-Dependent Modulation of Auditory-Evoked Responses in Cortex , 1999, The Journal of Neuroscience.

[22]  M. Stryker,et al.  Modulation of Visual Responses by Behavioral State in Mouse Visual Cortex , 2010, Neuron.

[23]  R. Yuste,et al.  Attractor dynamics of network UP states in the neocortex , 2003, Nature.

[24]  Patrick J. Mineault,et al.  Spatial clustering of tuning in mouse primary visual cortex , 2016, Nature Communications.

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

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

[27]  R. Yuste,et al.  Dynamics of Spontaneous Activity in Neocortical Slices , 2001, Neuron.

[28]  F. Helmchen,et al.  New angles on neuronal dendrites in vivo. , 2007, Journal of neurophysiology.

[29]  Takashi R Sato,et al.  Characterization and adaptive optical correction of aberrations during in vivo imaging in the mouse cortex , 2011, Proceedings of the National Academy of Sciences.

[30]  Rafael Yuste,et al.  Identification and Targeting of Cortical Ensembles , 2017, bioRxiv.

[31]  Amiram Grinvald,et al.  Iso-orientation domains in cat visual cortex are arranged in pinwheel-like patterns , 1991, Nature.

[32]  A. Gordus,et al.  Sensitive red protein calcium indicators for imaging neural activity , 2016, bioRxiv.

[33]  Bryan J MacLennan,et al.  Functional clustering of dendritic activity during decision-making , 2018, bioRxiv.

[34]  Sooyoung Chung,et al.  Functional imaging with cellular resolution reveals precise micro-architecture in visual cortex , 2005, Nature.

[35]  G. Buzsáki,et al.  Sequential structure of neocortical spontaneous activity in vivo , 2007, Proceedings of the National Academy of Sciences.

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

[37]  D. Hubel,et al.  Receptive fields, binocular interaction and functional architecture in the cat's visual cortex , 1962, The Journal of physiology.

[38]  T. Wilson,et al.  An optical technique for remote focusing in microscopy , 2008 .

[39]  Claire E McKellar,et al.  Rational design of a high-affinity, fast, red calcium indicator R-CaMP2 , 2014, Nature Methods.

[40]  Rainer W Friedrich,et al.  Remote z-scanning with a macroscopic voice coil motor for fast 3D multiphoton laser scanning microscopy , 2016, Biomedical optics express.

[41]  W. M. Keck,et al.  Highly Selective Receptive Fields in Mouse Visual Cortex , 2008, The Journal of Neuroscience.

[42]  R. Douglas,et al.  Neuronal circuits of the neocortex. , 2004, Annual review of neuroscience.

[43]  W. Denk,et al.  Dendritic spines as basic functional units of neuronal integration , 1995, Nature.

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

[45]  R. Reid,et al.  Specificity and randomness in the visual cortex , 2007, Current Opinion in Neurobiology.

[46]  Karel Svoboda,et al.  ScanImage: Flexible software for operating laser scanning microscopes , 2003, Biomedical engineering online.

[47]  R. Yuste.  From the neuron doctrine to neural networks , 2015, Nature Reviews Neuroscience.

[48]  Sreekanth H. Chalasani,et al.  Imaging neural activity in worms, flies and mice with improved GCaMP calcium indicators , 2009, Nature Methods.

[49]  Rafael Yuste,et al.  Control of postsynaptic Ca2+ influx in developing neocortex by excitatory and inhibitory neurotransmitters , 1991, Neuron.

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

[51]  Rafael Yuste,et al.  Imaging and Optically Manipulating Neuronal Ensembles. , 2017, Annual review of biophysics.

[52]  M. A. Smith,et al.  Spatial and Temporal Scales of Neuronal Correlation in Primary Visual Cortex , 2008, The Journal of Neuroscience.

[53]  Rafael Yuste,et al.  Endogenous Sequential Cortical Activity Evoked by Visual Stimuli , 2015, The Journal of Neuroscience.

[54]  Vítor Lopes-dos-Santos,et al.  Detecting cell assemblies in large neuronal populations , 2013, Journal of Neuroscience Methods.

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

[56]  Christine M Constantinople,et al.  Deep Cortical Layers Are Activated Directly by Thalamus , 2013, Science.

[57]  Olaf Sporns,et al.  Complex network measures of brain connectivity: Uses and interpretations , 2010, NeuroImage.

[58]  Jean-Loup Guillaume,et al.  Fast unfolding of communities in large networks , 2008, 0803.0476.

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

[60]  Geoffrey J. Goodhill,et al.  Spontaneous Activity in the Zebrafish Tectum Reorganizes over Development and Is Influenced by Visual Experience , 2017, Current Biology.

[61]  Alexander Attinger,et al.  Visuomotor Coupling Shapes the Functional Development of Mouse Visual Cortex , 2017, Cell.