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.
Published Web Locationhttps://doi.org/10.1038/nmeth.1552
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.