title: Fast volumetric calcium imaging across multiple cortical layers using sculpted light.
abstract: Although whole-organism calcium imaging in small and semi-transparent animals has been demonstrated, capturing the functional dynamics of large-scale neuronal circuits in awake behaving mammals at high speed and resolution has remained one of the main frontiers in systems neuroscience. Here we present a method based on light sculpting that enables unbiased single- and dual-plane high-speed (up to 160 Hz) calcium imaging as well as in vivo volumetric calcium imaging of a mouse cortical column (0.5 mm × 0.5 mm × 0.5 mm) at single-cell resolution and fast volume rates (3-6 Hz). We achieved this by tailoring the point-spread function of our microscope to the structures of interest while maximizing the signal-to-noise ratio using a home-built fiber laser amplifier with pulses that are synchronized to the imaging voxel speed. This enabled in vivo recording of calcium dynamics of several thousand neurons across cortical layers and in the hippocampus of awake behaving mice.
authors: Research Institute of Molecular Pathology, Vienna, Austria.|Max F. Perutz Laboratories Support GmbH, University of Vienna, Vienna, Austria.|Research Platform Quantum Phenomena &Nanoscale Biological Systems (QuNaBioS), University of Vienna, Vienna, Austria.|European Molecular Biology Laboratory, Heidelberg, Germany.
authors: Robert
authors: R
authors: Prevedel
authors: Photonics Institute, TU Wien, Vienna, Austria.|Center for Medical Physics and Biomedical Engineering, Medical University of Vienna, Vienna, Austria.
authors: Aart J
authors: AJ
authors: Verhoef
authors: Research Institute of Molecular Pathology, Vienna, Austria.
authors: Alejandro J
authors: AJ
authors: Pernía-Andrade
authors: Research Institute of Molecular Pathology, Vienna, Austria.|The Rockefeller University, New York, New York, USA.
authors: Siegfried
authors: S
authors: Weisenburger
authors: Department of Neurology and Psychiatry, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, California, USA.
authors: Ben S
authors: BS
authors: Huang
authors: Research Institute of Molecular Pathology, Vienna, Austria.|The Rockefeller University, New York, New York, USA.
authors: Tobias
authors: T
authors: Nöbauer
authors: Photonics Institute, TU Wien, Vienna, Austria.|Center for Medical Physics and Biomedical Engineering, Medical University of Vienna, Vienna, Austria.
authors: Alma
authors: A
authors: Fernández
authors: Research Institute of Molecular Pathology, Vienna, Austria.
authors: Jeroen E
authors: JE
authors: Delcour
authors: Department of Neurology and Psychiatry, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, California, USA.|West Los Angeles Virginia Medical Center, Los Angeles, California, USA.
authors: Peyman
authors: P
authors: Golshani
authors: Photonics Institute, TU Wien, Vienna, Austria.
authors: Andrius
authors: A
authors: Baltuska
authors: Research Institute of Molecular Pathology, Vienna, Austria.|Max F. Perutz Laboratories Support GmbH, University of Vienna, Vienna, Austria.|Research Platform Quantum Phenomena &Nanoscale Biological Systems (QuNaBioS), University of Vienna, Vienna, Austria.|The Rockefeller University, New York, New York, USA.
authors: Alipasha
authors: A
authors: Vaziri
mesh_terms: D000818:Animals; D001522:Behavior, Animal / Q000502:physiology; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling / Q000502:physiology*; D002540:Cerebral Cortex / Q000378:metabolism*; D006624:Hippocampus / Q000378:metabolism*; D051379:Mice; D018613:Microscopy, Confocal; D057054:Molecular Imaging / Q000379:methods*; D009474:Neurons / Q000378:metabolism*; D017785:Photons; D013997:Time Factors
chemical_list: D002118:Calcium