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

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Nature Methods. 2017 Jul;14(4):420 - 426. doi: 10.1038/nmeth.4226

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

Song A, Charles AS, Koay SA, Gauthier JL, Thiberge SY, Pillow JW, Tank DW

Note: Research in this publication was not performed at Janelia.

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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 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.

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

Note: Research in this publication was not performed at Janelia.

Abstract

Janelia Authors