Volumetric two-photon imaging of neurons using stereoscopy (vTwINS).

TitleVolumetric two-photon imaging of neurons using stereoscopy (vTwINS).
Publication TypeJournal Article
Year of Publication2017
AuthorsSong, A, Charles, AS, Koay, SAnn, Gauthier, JL, Thiberge, SY, Pillow, JW, Tank, DW
JournalNat Methods
Volume14
Issue4
Pagination420-426
Date Published2017 Apr
ISSN1548-7105
KeywordsAlgorithms, Animals, Calcium, Female, Hippocampus, Imaging, Three-Dimensional, Male, Mice, Transgenic, Microscopy, Confocal, Microscopy, Fluorescence, Multiphoton, Molecular Imaging, Neurons, Visual Cortex
Abstract

<p>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 uses 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 projections is proportional to depth in the volume. To demix the fluorescence time series of individual neurons, we introduce a modified orthogonal matching pursuit algorithm that also infers source locations within the 3D volume. We illustrated vTwINS by imaging neural population activity in the mouse primary visual cortex and hippocampus. Our results demonstrated 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.</p>

DOI10.1038/nmeth.4226
Alternate JournalNat. Methods
PubMed ID28319111
PubMed Central IDPMC5551981
Grant ListR01 MH083686 / MH / NIMH NIH HHS / United States
R34 MH083868 / MH / NIMH NIH HHS / United States
T32 MH065214 / MH / NIMH NIH HHS / United States
U01 NS090541 / NS / NINDS NIH HHS / United States