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Simultaneous two-photon calcium imaging at different depths with spatiotemporal multiplexing.

  • Author(s): Cheng, Adrian;
  • Gonçalves, J Tiago;
  • Golshani, Peyman;
  • Arisaka, Katsushi;
  • Portera-Cailliau, Carlos
  • et al.
Abstract

In vivo two-photon calcium imaging would benefit from the use of multiple excitation beams to increase scanning speed, signal-to-noise ratio and field of view or to image different axial planes simultaneously. Using spatiotemporal multiplexing we circumvented light-scattering ambiguity inherent to deep-tissue multifocal two-photon microscopy. We demonstrate calcium imaging at multiple axial planes in the intact mouse brain to monitor network activity of ensembles of cortical neurons in three spatial dimensions.

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