Fast 3D Imaging of Spine, Dendritic, and Neuronal Assemblies in Behaving Animals
Understanding neural computation requires methods such as 3D acousto-optical (AO) scanning that can simultaneously read out neural activity on both the somatic and dendritic scales. AO point scanning can increase measurement speed and signal-to-noise ratio (SNR) by several orders of magnitude, but h...
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| Published in | Neuron (Cambridge, Mass.) Vol. 92; no. 4; pp. 723 - 738 |
|---|---|
| Main Authors | , , , , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
United States
Elsevier Inc
23.11.2016
Elsevier Limited Cell Press |
| Subjects | |
| Online Access | Get full text |
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| Abstract | Understanding neural computation requires methods such as 3D acousto-optical (AO) scanning that can simultaneously read out neural activity on both the somatic and dendritic scales. AO point scanning can increase measurement speed and signal-to-noise ratio (SNR) by several orders of magnitude, but high optical resolution requires long point-to-point switching time, which limits imaging capability. Here we present a novel technology, 3D DRIFT AO scanning, which can extend each scanning point to small 3D lines, surfaces, or volume elements for flexible and fast imaging of complex structures simultaneously in multiple locations. Our method was demonstrated by fast 3D recording of over 150 dendritic spines with 3D lines, over 100 somata with squares and cubes, or multiple spiny dendritic segments with surface and volume elements, including in behaving animals. Finally, a 4-fold improvement in total excitation efficiency resulted in about 500 × 500 × 650 μm scanning volume with genetically encoded calcium indicators (GECIs).
•3D DRIFT AO scanning can extend each point to 3D lines, surfaces, or volume elements•Retained spatial information is used for post hoc movement correction of signals•Chessboard; snake; multi-cube; multi-3D line; and multi-layer, multi-frame scanning•Improved excitation efficiency for large 3D scanning volume with GECIs
3D DRIFT acousto-optical microscopy designed by Szalay et al. allows confined two-photon scanning of multiple 3D regions of interest. Activity of hundreds of neurons or multiple dendritic processes can be measured simultaneously in large cortical volumes of behaving animals. |
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| AbstractList | Understanding neural computation requires methods such as 3D acousto-optical (AO) scanning that can simultaneously read out neural activity on both the somatic and dendritic scales. AO point scanning can increase measurement speed and signal-to-noise ratio (SNR) by several orders of magnitude, but high optical resolution requires long point-to-point switching time, which limits imaging capability. Here we present a novel technology, 3D DRIFT AO scanning, which can extend each scanning point to small 3D lines, surfaces, or volume elements for flexible and fast imaging of complex structures simultaneously in multiple locations. Our method was demonstrated by fast 3D recording of over 150 dendritic spines with 3D lines, over 100 somata with squares and cubes, or multiple spiny dendritic segments with surface and volume elements, including in behaving animals. Finally, a 4-fold improvement in total excitation efficiency resulted in about 500 × 500 × 650 μm scanning volume with genetically encoded calcium indicators (GECIs). SummaryUnderstanding neural computation requires methods such as 3D acousto-optical (AO) scanning that can simultaneously read out neural activity on both the somatic and dendritic scales. AO point scanning can increase measurement speed and signal-to-noise ratio (SNR) by several orders of magnitude, but high optical resolution requires long point-to-point switching time, which limits imaging capability. Here we present a novel technology, 3D DRIFT AO scanning, which can extend each scanning point to small 3D lines, surfaces, or volume elements for flexible and fast imaging of complex structures simultaneously in multiple locations. Our method was demonstrated by fast 3D recording of over 150 dendritic spines with 3D lines, over 100 somata with squares and cubes, or multiple spiny dendritic segments with surface and volume elements, including in behaving animals. Finally, a 4-fold improvement in total excitation efficiency resulted in about 500 × 500 × 650 μm scanning volume with genetically encoded calcium indicators (GECIs). Understanding neural computation requires methods such as 3D acousto-optical (AO) scanning that can simultaneously read out neural activity on both the somatic and dendritic scales. AO point scanning can increase measurement speed and signal-to-noise ratio (SNR) by several orders of magnitude, but high optical resolution requires long point-to-point switching time, which limits imaging capability. Here we present a novel technology, 3D DRIFT AO scanning, which can extend each scanning point to small 3D lines, surfaces, or volume elements for flexible and fast imaging of complex structures simultaneously in multiple locations. Our method was demonstrated by fast 3D recording of over 150 dendritic spines with 3D lines, over 100 somata with squares and cubes, or multiple spiny dendritic segments with surface and volume elements, including in behaving animals. Finally, a 4-fold improvement in total excitation efficiency resulted in about 500 × 500 × 650 μm scanning volume with genetically encoded calcium indicators (GECIs). • 3D DRIFT AO scanning can extend each point to 3D lines, surfaces, or volume elements • Retained spatial information is used for post hoc movement correction of signals • Chessboard; snake; multi-cube; multi-3D line; and multi-layer, multi-frame scanning • Improved excitation efficiency for large 3D scanning volume with GECIs 3D DRIFT acousto-optical microscopy designed by Szalay et al. allows confined two-photon scanning of multiple 3D regions of interest. Activity of hundreds of neurons or multiple dendritic processes can be measured simultaneously in large cortical volumes of behaving animals. Understanding neural computation requires methods such as 3D acousto-optical (AO) scanning that can simultaneously read out neural activity on both the somatic and dendritic scales. AO point scanning can increase measurement speed and signal-to-noise ratio (SNR) by several orders of magnitude, but high optical resolution requires long point-to-point switching time, which limits imaging capability. Here we present a novel technology, 3D DRIFT AO scanning, which can extend each scanning point to small 3D lines, surfaces, or volume elements for flexible and fast imaging of complex structures simultaneously in multiple locations. Our method was demonstrated by fast 3D recording of over 150 dendritic spines with 3D lines, over 100 somata with squares and cubes, or multiple spiny dendritic segments with surface and volume elements, including in behaving animals. Finally, a 4-fold improvement in total excitation efficiency resulted in about 500 × 500 × 650 μm scanning volume with genetically encoded calcium indicators (GECIs). •3D DRIFT AO scanning can extend each point to 3D lines, surfaces, or volume elements•Retained spatial information is used for post hoc movement correction of signals•Chessboard; snake; multi-cube; multi-3D line; and multi-layer, multi-frame scanning•Improved excitation efficiency for large 3D scanning volume with GECIs 3D DRIFT acousto-optical microscopy designed by Szalay et al. allows confined two-photon scanning of multiple 3D regions of interest. Activity of hundreds of neurons or multiple dendritic processes can be measured simultaneously in large cortical volumes of behaving animals. |
| Author | Ócsai, Katalin Veress, Máté Szadai, Zoltán Katona, Gergely Fehér, András Judák, Linda Chiovini, Balázs Maák, Pál Juhász, Gábor Tompa, Tamás Szalay, Gergely Rózsa, Balázs |
| AuthorAffiliation | 1 Laboratory of 3D Functional Network and Dendritic Imaging, Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest 1083, Hungary 4 Department of Atomic Physics, Budapest University of Technology and Economics, Budapest 1111, Hungary 2 MTA-PPKE ITK-NAP B–2p Measurement Technology Group, Faculty of Information Technology, Pázmány Péter Catholic University, Budapest 1083, Hungary 3 Two-Photon Laboratory, Faculty of Information Technology, Pázmány Péter Catholic University, Budapest 1083, Hungary |
| AuthorAffiliation_xml | – name: 3 Two-Photon Laboratory, Faculty of Information Technology, Pázmány Péter Catholic University, Budapest 1083, Hungary – name: 1 Laboratory of 3D Functional Network and Dendritic Imaging, Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest 1083, Hungary – name: 2 MTA-PPKE ITK-NAP B–2p Measurement Technology Group, Faculty of Information Technology, Pázmány Péter Catholic University, Budapest 1083, Hungary – name: 4 Department of Atomic Physics, Budapest University of Technology and Economics, Budapest 1111, Hungary |
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| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/27773582$$D View this record in MEDLINE/PubMed |
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| Title | Fast 3D Imaging of Spine, Dendritic, and Neuronal Assemblies in Behaving Animals |
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