A large field of view two-photon mesoscope with subcellular resolution for in vivo imaging

Imaging is used to map activity across populations of neurons. Microscopes with cellular resolution have small (< 1 millimeter) fields of view and cannot simultaneously image activity distributed across multiple brain areas. Typical large field of view microscopes do not resolve single cells, especially in the axial dimension. We developed a 2-photon random access mesoscope (2p-RAM) that allows high-resolution imaging anywhere within a volume spanning multiple brain areas (Ø 5 mm × 1 mm cylinder). 2p-RAM resolution is near diffraction limited (lateral, 0.66 μm, axial 4.09 μm at the center; excitation wavelength = 970 nm; numerical aperture = 0.6) over a large range of excitation wavelengths. A fast threedimensional scanning system allows efficient sampling of neural activity in arbitrary regions of interest across the entire imaging volume. We illustrate the use of the 2p-RAM by imaging neural activity in multiple, non-contiguous brain areas in transgenic mice expressing protein calcium sensors.

[1]  Xunbin Wei,et al.  Investigation on the optimal wavelength for two-photon microscopy in brain tissue , 2018 .

[2]  Kaspar Podgorski,et al.  Brain heating induced by near infrared lasers during multi-photon microscopy , 2016, bioRxiv.

[3]  Kaspar Podgorski,et al.  Brain heating induced by near infrared lasers during multi-photon microscopy , 2016, bioRxiv.

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

[5]  Amy Hu,et al.  Sensitive red protein calcium indicators for imaging neural activity , 2016, bioRxiv.

[6]  G. McConnell,et al.  The Mesolens Project at the University of Strathclyde , 2016 .

[7]  Karel Svoboda,et al.  Neural coding in barrel cortex during whisker-guided locomotion , 2015, eLife.

[8]  Michael W. Kudenov,et al.  Wide field-of-view, multi-region two-photon imaging of neuronal activity in vivo , 2015, bioRxiv.

[9]  David Kleinfeld,et al.  Ultra-large field-of-view two-photon microscopy. , 2015, Optics express.

[10]  Tsai-Wen Chen,et al.  Comprehensive imaging of cortical networks , 2015, Current Opinion in Neurobiology.

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

[12]  Nathan C. Klapoetke,et al.  Transgenic Mice for Intersectional Targeting of Neural Sensors and Effectors with High Specificity and Performance , 2015, Neuron.

[13]  Nicholas A. Steinmetz,et al.  Diverse coupling of neurons to populations in sensory cortex , 2015, Nature.

[14]  Benjamin F. Grewe,et al.  Visualizing mammalian brain area interactions by dual-axis two-photon calcium imaging , 2015, 2015 Conference on Lasers and Electro-Optics (CLEO).

[15]  T. Murphy,et al.  Mesoscale Transcranial Spontaneous Activity Mapping in GCaMP3 Transgenic Mice Reveals Extensive Reciprocal Connections between Areas of Somatomotor Cortex , 2014, The Journal of Neuroscience.

[16]  Jeffrey N. Stirman,et al.  Wide field-of-view, twin-region two-photon imaging across extended cortical networks , 2014 .

[17]  Brenda C. Shields,et al.  Thy1-GCaMP6 Transgenic Mice for Neuronal Population Imaging In Vivo , 2014, PloS one.

[18]  Karel Svoboda,et al.  Natural Whisker-Guided Behavior by Head-Fixed Mice in Tactile Virtual Reality , 2014, The Journal of Neuroscience.

[19]  M. Stryker,et al.  A Cortical Circuit for Gain Control by Behavioral State , 2014, Cell.

[20]  E. Boyden,et al.  Simultaneous whole-animal 3D-imaging of neuronal activity using light-field microscopy , 2014, Nature Methods.

[21]  Zengcai V. Guo,et al.  Flow of Cortical Activity Underlying a Tactile Decision in Mice , 2014, Neuron.

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

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

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

[25]  Lacey J. Kitch,et al.  Long-term dynamics of CA1 hippocampal place codes , 2013, Nature Neuroscience.

[26]  James E. Fitzgerald,et al.  Photon shot noise limits on optical detection of neuronal spikes and estimation of spike timing. , 2013, Biophysical journal.

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

[28]  Jasper Akerboom,et al.  Optimization of a GCaMP Calcium Indicator for Neural Activity Imaging , 2012, The Journal of Neuroscience.

[29]  Lin Tian,et al.  Activity in motor-sensory projections reveals distributed coding in somatosensation , 2012, Nature.

[30]  J. Simon Wiegert,et al.  Multiple dynamic representations in the motor cortex during sensorimotor learning , 2012, Nature.

[31]  Christine Grienberger,et al.  Imaging Calcium in Neurons , 2012, Neuron.

[32]  Christopher D. Harvey,et al.  Choice-specific sequences in parietal cortex during a virtual-navigation decision task , 2012, Nature.

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

[34]  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.

[35]  Yongxin Zhao,et al.  An Expanded Palette of Genetically Encoded Ca2+ Indicators , 2011, Science.

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

[37]  R. Reid,et al.  Broadly Tuned Response Properties of Diverse Inhibitory Neuron Subtypes in Mouse Visual Cortex , 2010, Neuron.

[38]  Takeharu Nagai,et al.  Spontaneous network activity visualized by ultrasensitive Ca2+ indicators, yellow Cameleon-Nano , 2010, Nature Methods.

[39]  R. Romo,et al.  Decoding a Perceptual Decision Process across Cortex , 2010, Neuron.

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

[41]  Aberration effects on femtosecond pulses generated by nonideal achromatic doublets. , 2009, Applied optics.

[42]  K. Svoboda,et al.  Long-term, high-resolution imaging in the mouse neocortex through a chronic cranial window , 2009, Nature Protocols.

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

[44]  T. Holy,et al.  Fast Three-Dimensional Fluorescence Imaging of Activity in Neural Populations by Objective-Coupled Planar Illumination Microscopy , 2008, Neuron.

[45]  Byron M. Yu,et al.  Techniques for extracting single-trial activity patterns from large-scale neural recordings , 2007, Current Opinion in Neurobiology.

[46]  K. Svoboda,et al.  The Functional Microarchitecture of the Mouse Barrel Cortex , 2007, Neuroscience Research.

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

[48]  Rick Trebino,et al.  Extremely simple single-prism ultrashort- pulse compressor. , 2006, Optics express.

[49]  P. Saggau,et al.  Fast three-dimensional laser scanning scheme using acousto-optic deflectors. , 2005, Journal of biomedical optics.

[50]  A. Miyawaki,et al.  Expanded dynamic range of fluorescent indicators for Ca(2+) by circularly permuted yellow fluorescent proteins. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[51]  Alan Peters,et al.  THE SMALL PYRAMIDAL NEURON OF THE RAT CEREBRAL CORTEX , 1968, Zeitschrift für Zellforschung und Mikroskopische Anatomie.

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

[53]  C. Stosiek,et al.  In vivo two-photon calcium imaging of neuronal networks , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[54]  K. Svoboda,et al.  Long-term in vivo imaging of experience-dependent synaptic plasticity in adult cortex , 2002, Nature.

[55]  Jerome Mertz,et al.  Two-photon microscopy in brain tissue: parameters influencing the imaging depth , 2001, Journal of Neuroscience Methods.

[56]  G. Feng,et al.  Imaging Neuronal Subsets in Transgenic Mice Expressing Multiple Spectral Variants of GFP , 2000, Neuron.

[57]  K. Svoboda,et al.  Two-photon imaging in living brain slices. , 1999, Methods.

[58]  M H Ellisman,et al.  Video-rate scanning two-photon excitation fluorescence microscopy and ratio imaging with cameleons. , 1999, Biophysical journal.

[59]  K. Svoboda,et al.  Photon Upmanship: Why Multiphoton Imaging Is More than a Gimmick , 1997, Neuron.

[60]  D. Kleinfeld,et al.  Anatomical and functional imaging of neurons using 2-photon laser scanning microscopy , 1994, Journal of Neuroscience Methods.

[61]  J. Pawley,et al.  Handbook of Biological Confocal Microscopy , 1990, Springer US.

[62]  A. Peters,et al.  The small pyramidal neuron of the rat cerebral cortex. The perikaryon, dendrites and spines. , 1970, The American journal of anatomy.

[63]  Alan Peters,et al.  THE SMALL PYRAMIDAL NEURON OF THE RAT CEREBRAL CORTEX The Axon Hillock and Initial Segment , 1968 .

[64]  Sanford L. Palay,et al.  THE AXON HILLOCK AND THE INITIAL SEGMENT , 1968, The Journal of cell biology.

[65]  Emil Wolf,et al.  Principles of Optics: Contents , 1999 .