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Volumetric Two-photon Imaging of Neurons Using Stereoscopy (vTwINS)

Alexander Song, View ORCID ProfileAdam S. Charles, Sue Ann Koay, Jeff L. Gauthier, Stephan Y. Thiberge, Jonathan W. Pillow, David W. Tank
doi: https://doi.org/10.1101/073742
Alexander Song
Princeton University
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Adam S. Charles
Princeton University
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  • ORCID record for Adam S. Charles
Sue Ann Koay
Princeton University
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Jeff L. Gauthier
Princeton University
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Stephan Y. Thiberge
Princeton University
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Jonathan W. Pillow
Princeton University
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David W. Tank
Princeton University
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  • For correspondence: dwtank@princeton.edu
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Abstract

Two-photon laser scanning microscopy of calcium dynamics using fluorescent indicators is a widely used imaging method for large scale recording of neural activity in vivo. Here we introduce volumetric Two-photon Imaging of Neurons using Stereoscopy (vTwINS), a volumetric calcium imaging method that employs an elongated, V-shaped point spread function to image a 3D brain volume. Single neurons project to spatially displaced "image pairs" in the resulting 2D image, and the separation distance between images is proportional to depth in the volume. To demix the fluorescence time series of individual neurons, we introduce a novel orthogonal matching pursuit algorithm that also infers source locations within the 3D volume. We illustrate vTwINS by imaging neural population activity in mouse primary visual cortex and hippocampus. Our results demonstrate that vTwINS provides an effective method for volumetric two-photon calcium imaging that increases the number of neurons recorded while maintaining a high frame-rate.

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Posted September 06, 2016.
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Volumetric Two-photon Imaging of Neurons Using Stereoscopy (vTwINS)
Alexander Song, Adam S. Charles, Sue Ann Koay, Jeff L. Gauthier, Stephan Y. Thiberge, Jonathan W. Pillow, David W. Tank
bioRxiv 073742; doi: https://doi.org/10.1101/073742
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Volumetric Two-photon Imaging of Neurons Using Stereoscopy (vTwINS)
Alexander Song, Adam S. Charles, Sue Ann Koay, Jeff L. Gauthier, Stephan Y. Thiberge, Jonathan W. Pillow, David W. Tank
bioRxiv 073742; doi: https://doi.org/10.1101/073742

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