Three-dimensional imaging and photostimulation by remote-focusing and holographic light patterning
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 digita...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 2011-12, Vol.108 (49), p.19504-19509 |
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| container_title | Proceedings of the National Academy of Sciences - PNAS |
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| creator | Anselmi, Francesca Ventalon, Cathie Bègue, Aurélien Ogden, David Emiliani, Valentina |
| description | 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. |
| doi_str_mv | 10.1073/pnas.1109111108 |
| format | Article |
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| ispartof | Proceedings of the National Academy of Sciences - PNAS, 2011-12, Vol.108 (49), p.19504-19509 |
| issn | 0027-8424 1091-6490 |
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| subjects | Algorithms Animals Biological Sciences Brain CA1 Region, Hippocampal - cytology CA1 Region, Hippocampal - metabolism Calcium Calcium - metabolism Dendrites Dendrites - metabolism Dendrites - physiology Electric currents Glutamic Acid - metabolism Hippocampus - anatomy & histology Hippocampus - cytology Hippocampus - metabolism Holography Holography - instrumentation Holography - methods image analysis Imaging Imaging, Three-Dimensional - instrumentation Imaging, Three-Dimensional - methods Ion Channel Gating - drug effects Lasers Light Mice Mice, Inbred Strains Microscopes Microscopy Microscopy, Fluorescence Neurons Optical focus Patch-Clamp Techniques Photoactivation Photolysis Physical Sciences Point sources Pyramidal Cells - metabolism Pyramidal Cells - physiology Sodium Channel Blockers - pharmacology Sodium Channels - metabolism Tetrodotoxin - pharmacology Three dimensional imaging |
| title | Three-dimensional imaging and photostimulation by remote-focusing and holographic light patterning |
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