Title | Volumetric two-photon imaging of neurons using stereoscopy (vTwINS). |
Publication Type | Journal Article |
Year of Publication | 2017 |
Authors | Song, A, Charles, AS, Koay, SAnn, Gauthier, JL, Thiberge, SY, Pillow, JW, Tank, DW |
Journal | Nat Methods |
Volume | 14 |
Issue | 4 |
Pagination | 420-426 |
Date Published | 2017 Apr |
ISSN | 1548-7105 |
Keywords | Algorithms, 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> |
DOI | 10.1038/nmeth.4226 |
Alternate Journal | Nat Methods |
PubMed ID | 28319111 |
PubMed Central ID | PMC5551981 |
Grant List | R01 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 |