Sustained deep-tissue voltage recording using a fast indicator evolved for two-photon microscopy

Cell. 2022 Sep 1;185(18):3408-3425.e29. doi: 10.1016/j.cell.2022.07.013. Epub 2022 Aug 18.

Abstract

Genetically encoded voltage indicators are emerging tools for monitoring voltage dynamics with cell-type specificity. However, current indicators enable a narrow range of applications due to poor performance under two-photon microscopy, a method of choice for deep-tissue recording. To improve indicators, we developed a multiparameter high-throughput platform to optimize voltage indicators for two-photon microscopy. Using this system, we identified JEDI-2P, an indicator that is faster, brighter, and more sensitive and photostable than its predecessors. We demonstrate that JEDI-2P can report light-evoked responses in axonal termini of Drosophila interneurons and the dendrites and somata of amacrine cells of isolated mouse retina. JEDI-2P can also optically record the voltage dynamics of individual cortical neurons in awake behaving mice for more than 30 min using both resonant-scanning and ULoVE random-access microscopy. Finally, ULoVE recording of JEDI-2P can robustly detect spikes at depths exceeding 400 μm and report voltage correlations in pairs of neurons.

Keywords: GEVI; JEDI-2P; fly vision; genetically encoded voltage indicator; high-throughput screening; pairwise voltage correlations; random-access microscopy; starburst amacrine cells; two-photon fluorescence microscopy; voltage imaging.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Interneurons
  • Mice
  • Microscopy* / methods
  • Neurons* / physiology
  • Photons
  • Wakefulness