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

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

Research output: Contribution to journalArticlepeer-review

96 Scopus citations


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 modifed 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.

Original languageEnglish (US)
Pages (from-to)420-426
Number of pages7
JournalNature Methods
Issue number4
StatePublished - Mar 20 2017

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Biochemistry
  • Biotechnology
  • Cell Biology


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