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Proceedings Paper

In vivo three-photon activity imaging of GCaMP6-labeled neurons in deep cortex and the hippocampus of the mouse brain
Author(s): Tianyu Wang; Dimitre G. Ouzounov; Mengran Wang; Danielle Feng; Jean C. Cruz-Hernandez; Jacob Reimer; Andreas Tolias; Nozomi Nishimura; Chris Xu
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Paper Abstract

We demonstrate that three-photon microscopy (3PM) with 1300-nm excitation enables functional imaging of GCaMP6s labeled neurons beyond the depth limit of two-photon microscopy (2PM) with 920-nm excitation. We quantitatively compared 2PM and 3PM imaging of calcium indicator GCaMP6s by measuring correlation between activity traces, absolute signal level, excitation attenuation with depth, and signal-to-background ratio (SBR). Compared to 2PM imaging of GCaMP6s-labeled neurons, 3PM imaging has increasingly larger advantages in signal strength and SBR as the imaging depth increases in densely labeled mouse brain, given the same pulse energy, pulse width, and repetition rate at the sample surface. For example, 3PM has comparable signal strength as 2PM and up to two orders of magnitude higher SBR as 2PM in mouse cortex around 700-800um. We also demonstrate 3PM activity recording of 150 neurons in the hippocampal stratum pyramidale (SP) at 1mm depth, which is inaccessible to non-invasive 2PM imaging. Our work establishes 3PM as a powerful tool for calcium imaging at the depth beyond 2PM limits.

Paper Details

Date Published: 2 March 2017
PDF: 5 pages
Proc. SPIE 10069, Multiphoton Microscopy in the Biomedical Sciences XVII, 100692L (2 March 2017); doi: 10.1117/12.2251220
Show Author Affiliations
Tianyu Wang, Cornell Univ. (United States)
Dimitre G. Ouzounov, Cornell Univ. (United States)
Mengran Wang, Cornell Univ. (United States)
Danielle Feng, Cornell Univ. (United States)
Jean C. Cruz-Hernandez, Cornell Univ. (United States)
Jacob Reimer, Baylor College of Medicine (United States)
Andreas Tolias, Baylor College of Medicine (United States)
Nozomi Nishimura, Cornell Univ. (United States)
Chris Xu, Cornell Univ. (United States)

Published in SPIE Proceedings Vol. 10069:
Multiphoton Microscopy in the Biomedical Sciences XVII
Ammasi Periasamy; Peter T. C. So; Karsten König; Xiaoliang S. Xie, Editor(s)

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