Journal
NATURE METHODS
Volume 17, Issue 1, Pages 107-+Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/s41592-019-0625-2
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Funding
- NIH [MH099045, EY022951, NS094358, EY026878, MH111424, EY029581, GM007205, NS007224, EY028869]
- William Ziegler III family
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Simultaneous two-photon microscopic and one-photon mesoscopic imaging of calcium signals enables correlation of local cellular and brain-wide network activity. Spontaneous and sensory-evoked activity propagates across varying spatial scales in the mammalian cortex, but technical challenges have limited conceptual links between the function of local neuronal circuits and brain-wide network dynamics. We present a method for simultaneous cellular-resolution two-photon calcium imaging of a local microcircuit and mesoscopic widefield calcium imaging of the entire cortical mantle in awake mice. Our multi-scale approach involves a microscope with an orthogonal axis design where the mesoscopic objective is oriented above the brain and the two-photon objective is oriented horizontally, with imaging performed through a microprism. We also introduce a viral transduction method for robust and widespread gene delivery in the mouse brain. These approaches allow us to identify the behavioral state-dependent functional connectivity of pyramidal neurons and vasoactive intestinal peptide-expressing interneurons with long-range cortical networks. Our imaging system provides a powerful strategy for investigating cortical architecture across a wide range of spatial scales.
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