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Cell. 2019 May 2;177(4):1050-1066.e14. doi: 10.1016/j.cell.2019.03.011. Epub 2019 Apr 11.

Volumetric Ca2+ Imaging in the Mouse Brain Using Hybrid Multiplexed Sculpted Light Microscopy.

Author information

1
Laboratory of Neurotechnology and Biophysics, The Rockefeller University, New York, NY, USA.
2
Department of Neuroscience, Columbia University, New York, NY, USA; Mortimer B. Zuckerman Mind Brain Behavior Institute, Columbia University, New York, NY, USA.
3
Allen Institute for Brain Science, Seattle, WA, USA.
4
Department of Neuroscience, Columbia University, New York, NY, USA; The Kavli Institute for Brain Science, Columbia University, New York, NY, USA; Mortimer B. Zuckerman Mind Brain Behavior Institute, Columbia University, New York, NY, USA.
5
Laboratory of Neurotechnology and Biophysics, The Rockefeller University, New York, NY, USA; Research Institute of Molecular Pathology, Vienna, Austria; The Kavli Neural Systems Institute, The Rockefeller University, New York, NY, USA. Electronic address: vaziri@rockefeller.edu.

Abstract

Calcium imaging using two-photon scanning microscopy has become an essential tool in neuroscience. However, in its typical implementation, the tradeoffs between fields of view, acquisition speeds, and depth restrictions in scattering brain tissue pose severe limitations. Here, using an integrated systems-wide optimization approach combined with multiple technical innovations, we introduce a new design paradigm for optical microscopy based on maximizing biological information while maintaining the fidelity of obtained neuron signals. Our modular design utilizes hybrid multi-photon acquisition and allows volumetric recording of neuroactivity at single-cell resolution within up to 1 × 1 × 1.22 mm volumes at up to 17 Hz in awake behaving mice. We establish the capabilities and potential of the different configurations of our imaging system at depth and across brain regions by applying it to in vivo recording of up to 12,000 neurons in mouse auditory cortex, posterior parietal cortex, and hippocampus.

KEYWORDS:

2-photon; 3-photon; Ca(2+) imaging; circuit dynamics; cortical network; high-speed; light sculpting; microscopy; systems neuroscience; volumetric

PMID:
30982596
DOI:
10.1016/j.cell.2019.03.011

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