Widefield imaging of rapid pan-cortical voltage dynamics with an indicator evolved for one-photon microscopy.

Authors

Xiaoyu Lu
Yunmiao Wang
Zhuohe Liu
Yueyang Gou
Dieter Jaeger
François St-Pierre

Publication Date

10-12-2023

Journal

Nature Communications

DOI

10.1038/s41467-023-41975-3

PMID

37828037

PMCID

PMC10570354

PubMedCentral® Posted Date

10-12-2023

PubMedCentral® Full Text Version

Post-print

Published Open-Access

yes

Keywords

Mice, Animals, Microscopy, Neurons, Photons, Brain, Photic Stimulation, Optical imaging, Neuroscience, Sensors and probes, High-throughput screening

Abstract

Widefield imaging with genetically encoded voltage indicators (GEVIs) is a promising approach for understanding the role of large cortical networks in the neural coding of behavior. However, the limited performance of current GEVIs restricts their deployment for single-trial imaging of rapid neuronal voltage dynamics. Here, we developed a high-throughput platform to screen for GEVIs that combine fast kinetics with high brightness, sensitivity, and photostability under widefield one-photon illumination. Rounds of directed evolution produced JEDI-1P, a green-emitting fluorescent indicator with enhanced performance across all metrics. Next, we optimized a neonatal intracerebroventricular delivery method to achieve cost-effective and wide-spread JEDI-1P expression in mice. We also developed an approach to correct optical measurements from hemodynamic and motion artifacts effectively. Finally, we achieved stable brain-wide voltage imaging and successfully tracked gamma-frequency whisker and visual stimulations in awake mice in single trials, opening the door to investigating the role of high-frequency signals in brain computations.

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