Neuron. 2016 Nov 23;92(4):723-738. doi: 10.1016/j.neuron.2016.10.002.

Fast 3D Imaging of Spine, Dendritic, and Neuronal Assemblies in Behaving Animals.

Szalay G1, Judák L1, Katona G2, Ócsai K3, Juhász G2, Veress M4, Szadai Z5, Fehér A4, Tompa T5, Chiovini B5, Maák P4, Rózsa B6.

Author information

1 Laboratory of 3D Functional Network and Dendritic Imaging, Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest 1083, Hungary.
2 Laboratory of 3D Functional Network and Dendritic Imaging, Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest 1083, Hungary; MTA-PPKE ITK-NAP B-2p Measurement Technology Group, Faculty of Information Technology, Pázmány Péter Catholic University, Budapest 1083, Hungary.
3 MTA-PPKE ITK-NAP B-2p Measurement Technology Group, Faculty of Information Technology, Pázmány Péter Catholic University, Budapest 1083, Hungary.
4 Department of Atomic Physics, Budapest University of Technology and Economics, Budapest 1111, Hungary.
5 Laboratory of 3D Functional Network and Dendritic Imaging, Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest 1083, Hungary; Two-Photon Laboratory, Faculty of Information Technology, Pázmány Péter Catholic University, Budapest 1083, Hungary.
6 Laboratory of 3D Functional Network and Dendritic Imaging, Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest 1083, Hungary; Two-Photon Laboratory, Faculty of Information Technology, Pázmány Péter Catholic University, Budapest 1083, Hungary. Electronic address: rozsabal@koki.hu.
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).

Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

PMID: 27773582; PMCID: PMC5167293

 

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