RT Journal Article
SR Electronic
T1 Detecting rapid pan-cortical voltage dynamics in vivo with a brighter and faster voltage indicator
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.08.29.505018
DO 10.1101/2022.08.29.505018
A1 Xiaoyu Lu
A1 Yunmiao Wang
A1 Zhuohe Liu
A1 Yueyang Gou
A1 Dieter Jaeger
A1 François St-Pierre
YR 2022
UL http://biorxiv.org/content/early/2022/08/31/2022.08.29.505018.abstract
AB 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 slow kinetics of current GEVIs limit 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 whose performance is improved for 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 effectively correct optical measurements from hemodynamic and motion artifacts. 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.Competing Interest StatementFSP holds a US patent for a voltage sensor design (patent #US9606100 B2).