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Neuron. 2016 Jan 20;89(2):269-84. doi: 10.1016/j.neuron.2015.12.012. Epub 2016 Jan 7.

Simultaneous Multi-plane Imaging of Neural Circuits.

Author information

1
Neurotechnology Center, Department of Biological Sciences, Columbia University, New York, NY 10027, USA. Electronic address: wy2221@columbia.edu.
2
Neurotechnology Center, Department of Biological Sciences, Columbia University, New York, NY 10027, USA.
3
Center for Computational Biology, Simons Foundation, New York, NY 10010, USA; Department of Statistics, Center for Theoretical Neuroscience, and Grossman Center for the Statistics of Mind, Columbia University, New York, NY 10027, USA.
4
Neurotechnology Center, Department of Biological Sciences, Columbia University, New York, NY 10027, USA; Department of Statistics, Center for Theoretical Neuroscience, and Grossman Center for the Statistics of Mind, Columbia University, New York, NY 10027, USA.

Abstract

Recording the activity of large populations of neurons is an important step toward understanding the emergent function of neural circuits. Here we present a simple holographic method to simultaneously perform two-photon calcium imaging of neuronal populations across multiple areas and layers of mouse cortex in vivo. We use prior knowledge of neuronal locations, activity sparsity, and a constrained nonnegative matrix factorization algorithm to extract signals from neurons imaged simultaneously and located in different focal planes or fields of view. Our laser multiplexing approach is simple and fast, and could be used as a general method to image the activity of neural circuits in three dimensions across multiple areas in the brain.

PMID:
26774159
PMCID:
PMC4724224
DOI:
10.1016/j.neuron.2015.12.012
[Indexed for MEDLINE]
Free PMC Article

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