title: Three-dimensional imaging and photostimulation by remote-focusing and holographic light patterning.
abstract: Access to three-dimensional structures in the brain is fundamental to probe signal processing at multiple levels, from integration of synaptic inputs to network activity mapping. Here, we present an optical method for independent three-dimensional photoactivation and imaging by combination of digital holography with remote-focusing. We experimentally demonstrate compensation of spherical aberration for out-of-focus imaging in a range of at least 300 μm, as well as scanless imaging along oblique planes. We apply this method to perform functional imaging along tilted dendrites of hippocampal pyramidal neurons in brain slices, after photostimulation by multiple spots glutamate uncaging. By bringing extended portions of tilted dendrites simultaneously in-focus, we monitor the spatial extent of dendritic calcium signals, showing a shift from a widespread to a spatially confined response upon blockage of voltage-gated Na(+) channels.
authors.affiliation: Wavefront-Engineering Microscopy Group, Neurophysiology and New Microscopies Laboratory, Centre National de la Recherche Scientifique UMR8154, Institut National de la Santé et de la Recherche Médicale U603, Université Paris Descartes, 45 Rue des Saints Pères, F-75006 Paris, France.
authors.forename: Francesca
authors.initials: F
authors.lastname: Anselmi
authors.forename: Cathie
authors.initials: C
authors.lastname: Ventalon
authors.forename: Aurélien
authors.initials: A
authors.lastname: Bègue
authors.forename: David
authors.initials: D
authors.lastname: Ogden
authors.forename: Valentina
authors.initials: V
authors.lastname: Emiliani
mesh_terms: D000465:Algorithms; D000818:Animals; D056547:CA1 Region, Hippocampal / Q000166:cytology / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D003712:Dendrites / Q000378:metabolism / Q000502:physiology*; D018698:Glutamic Acid / Q000378:metabolism; D006624:Hippocampus / Q000033:anatomy & histology / Q000166:cytology / Q000378:metabolism; D006696:Holography / Q000295:instrumentation / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000295:instrumentation / Q000379:methods*; D015640:Ion Channel Gating / Q000187:drug effects; D008027:Light; D051379:Mice; D008815:Mice, Inbred Strains; D008856:Microscopy, Fluorescence; D018408:Patch-Clamp Techniques; D017966:Pyramidal Cells / Q000378:metabolism / Q000502:physiology; D026941:Sodium Channel Blockers / Q000494:pharmacology; D015222:Sodium Channels / Q000378:metabolism; D013779:Tetrodotoxin / Q000494:pharmacology
chemical_list: D026941:Sodium Channel Blockers; D015222:Sodium Channels; D018698:Glutamic Acid; D013779:Tetrodotoxin; D002118:Calcium
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