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Nat Biotechnol. 2016 Aug;34(8):857-62. doi: 10.1038/nbt.3594. Epub 2016 Jun 27.

Wide field-of-view, multi-region, two-photon imaging of neuronal activity in the mammalian brain.

Author information

1
Neuroscience Center, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.
2
Carolina Institute for Developmental Disabilities, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.
3
Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.
4
Department of Electrical and Computer Engineering, North Carolina State University, Raleigh, North Carolina, USA.
5
Department of Cell Biology and Physiology, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.
6
Neurobiology Curriculum, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.

Abstract

Two-photon calcium imaging provides an optical readout of neuronal activity in populations of neurons with subcellular resolution. However, conventional two-photon imaging systems are limited in their field of view to ∼1 mm(2), precluding the visualization of multiple cortical areas simultaneously. Here, we demonstrate a two-photon microscope with an expanded field of view (>9.5 mm(2)) for rapidly reconfigurable simultaneous scanning of widely separated populations of neurons. We custom designed and assembled an optimized scan engine, objective, and two independently positionable, temporally multiplexed excitation pathways. We used this new microscope to measure activity correlations between two cortical visual areas in mice during visual processing.

PMID:
27347754
PMCID:
PMC4980167
DOI:
10.1038/nbt.3594
[Indexed for MEDLINE]
Free PMC Article

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