Journal Article

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

Gauthier, Jeff L, Thiberge, Stephan Y, Charles, Adam S, Tank, David W, Song, Alexander, Pillow, Jonathan W, Koay, Sue Ann

2017

Overview

vTwINS enables high-speed volumetric calcium imaging via a V-shaped point spread function and a dedicated data-processing algorithm. Song et al . apply this strategy to image population activity in the mouse visual cortex and hippocampus. 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.

• Publisher: Nature Publishing Group US,Nature Publishing Group
• Subject: 631/1647/245/2226 / 631/1647/328/2057 / 631/1647/328/2235 / 631/1647/334/1874/345 / 631/378 / Algorithms / Animals / Bioinformatics / Biological Microscopy / Biological Techniques / Biomedical Engineering/Biotechnology / Brain / Calcium / Calcium - analysis / Calcium - metabolism / Calcium imaging / Calcium signalling / Confocal microscopy / Elongation / Female / Fluorescent indicators / Hippocampus - cytology / Hippocampus - physiology / Imaging / Imaging, Three-Dimensional - methods / In vivo methods and tests / Life Sciences / Male / Mice, Transgenic / Microscopy, Confocal - instrumentation / Microscopy, Confocal - methods / Microscopy, Fluorescence, Multiphoton - instrumentation / Microscopy, Fluorescence, Multiphoton - methods / Molecular Imaging - methods / Neuroimaging / Neurons / Neurons - physiology / Point spread functions / Proteomics / Recording / Scanning microscopy / Stereoscopy / Visual cortex / Visual Cortex - cytology / Visual Cortex - physiology