Presentation Abstract

Program#/Poster#: 871.24/MMM20
Presentation Title: Synchronous 3-D imaging of large neural populations with single-neuron resolution at video rates In vitro and In vivo with light field microscopy
Location: Halls B-H
Presentation time: Wednesday, Nov 13, 2013, 4:00 PM - 5:00 PM
Topic: ++G.04.a. Optical methods
Authors: *L. GROSENICK1,2,3,4, M. BROXTON5, C. K. KIM1,3, C. LISTON1, S. YANG6, A. ANDALMAN1,4, N. COHEN6, L. LEUNG7, B. POOLE5, J. VOGELSTEIN10,11, T. ANDERSON8, Z. ZHANG5, O. YIZHAR12, B. GRONE7, C. RAMAKRISHNAN1, A. MUTO13, K. KAWAKAMI13, P. MOURRAIN7, S. J. SMITH8, P. SUPPES9, M. LEVOY5,6, K. DEISSEROTH1,4,7,14;
1Bioengineering, 2Ctr. for Mind, Brain, and Computation, 3Neurosciences Program, 4CNC Program, 5Computer Sci., 6Electrical Engin., 7Psychiatry and Behavioral Sci., 8Cell. and Mol. Physiol., 9Philosophy, Stanford Univ., Stanford, CA; 10Dept. of Statistical Sci., 11Inst. for Brain Sci., Duke Univ., Durham, NC; 12Neurobio., Weizmann Inst. of Sci., Rehovot, Israel; 13Div. of Mol. and Developmental Biol., Grad. Univ. for Advanced Studies (SOKENDAI), Mishima, Japan; 14HHMI, Chevy Chase, MD
Abstract: Behaviorally-relevant aspects of neural population dynamics can unfold on a timescale of milliseconds. Although recent developments in optical sensing of neural activity approach this temporal resolution, imaging fast circuit dynamics over the large populations of neurons that generate behavior will require methods capable of synchronously capturing large 3-D volumes at high frame rates with single neuron resolution. Recent years have seen the introduction of several promising volumetric imaging approaches (Abrahamsson et al., 2013 , Gobel et al., 2007, Holekamp et al., 2008, Niesner et al., 2007, Reddy et al., 2008, Tomer et al, 2012). However, each of these requires a sequential capture of the volumes over time or a sharp division of the camera sensor into regions corresponding to each axial plane. This restricts the utility of these approaches for rapid functional imaging of many neurons. We demonstrate for the first time that light field microscopy (Levoy et al., 2006) can be used in biology, here to acquire synchronous 3-D images of genetically-encoded calcium indicator (GECI) activity in vitro and in vivo with single-neuron resolution at speeds limited only by the maximum camera frame rate (100 fps with current sCMOS cameras). Although diffraction imposes a trade-off between lateral and axial resolution, we show that a novel approach to light field deconvolution yields a 2-4 fold improvement in lateral resolution over previously reported light field micrographs. The large volume of functional data (up to 1.6 mm x 1.4 mm x 0.5 mm with a 10x objective, 0.8 mm x 0.7 mm x 0.4 mm with a 20x objective) covered by this method allows whole-brain imaging of transgenic larval zebrafish (5-14 dpf) expressing GCaMP7a (Muto et al., 2013) under a pan-neuronal promoter. This approach is compatible with head-fixed, visually-mediated behavior in a simple virtual environment, and the majority of the zebrafish brain can be imaged synchronously with single neurons resolvable over an appreciable axial range. Without moving the zebrafish, it is possible to collect registered 3-D confocal or two-photon stacks to recover additional structural information. Using virally-transduced, genetically-encoded indicator GCaMP6f in mice, activity in single neurons can be resolved at up to 100 fps, allowing 10 ms temporal resolution over a volume covering all six layers in an acute cortical slice preparation or areas CA1-CA3 of hippocampus imaged in an awake, behaving mouse on a track ball (Dombeck, 2010). Finally, we show that using factor-analytic and temporal-deconvolution algorithms, these data can be used to estimate the number of spikes occurring per camera frame for each neural source.
Disclosures:  L. Grosenick: None. M. Broxton: None. C.K. Kim: None. C. Liston: None. S. Yang: None. A. Andalman: None. N. Cohen: None. L. Leung: None. B. Poole: None. J. Vogelstein: None. T. Anderson: None. Z. Zhang: None. O. Yizhar: None. B. Grone: None. C. Ramakrishnan: None. P. Mourrain: None. S.J. Smith: None. P. Suppes: None. M. Levoy: None. K. Deisseroth: None. A. Muto: None. K. Kawakami: None.
Keyword(s): IMAGING
CALCIUM IMAGING
IN VIVO
Support: HHMI
NIH
NSF
DARPA
SFARI




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