A red-emitting, genetically encoded indicator for two-photon voltage recording in vivo
Abstract
Genetically encoded voltage indicators (GEVIs) enable minimally invasive, cell-type-specific optical measurements of neuronal membrane potential with millisecond temporal resolution. Red-shifted GEVIs are especially advantageous because they permit spectral multiplexing with complementary sensors and enable all-optical circuit interrogation in combination with blue-light-activated opsins. Despite these advantages, existing red GEVIs remain poorly suited for in vivo use due to limited performance under two-photon (2P) excitation, the predominant modality for deep-tissue imaging. Here, we introduce VADER1, a red GEVI that overcomes this limitation and enables reliable spike detection in vivo under 2P illumination. Under 2P excitation, VADER1 supports extended voltage imaging with both random-access and resonant-scanning microscopy, enables recordings from neurons as deep as cortical layer 5, and allows dual-color imaging with calcium indicators. By filling a critical spectral gap, VADER1 enables integrated optical measurements of fast electrical activity alongside other neural signals and establishes a foundation for two-photon all-optical electrophysiology.
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BibTeX
@misc{yang2026redemitting,
title = {{A red-emitting, genetically encoded indicator for two-photon voltage recording in vivo}},
author = {Shuyuan Yang and Alex James Mcdonald and Xiaoyu Lu and Vincent Villette and Noura Hakam and Mario Galdamez and Tanmayee Torne-Srivas and Gregory Foran and Xiaoyu Dong and Matthew Shorey and Nima Bavili and Shujuan Lai and Zhuohe Liu and Haixin Liu and Tom Maslianitsyna and Rémi Fournel and Emiliano Ronzitti and Jun Zhu and Ryan Gregory Natan and Jonathan Bradley and Daniela Gaspar Santos and Yu-Yau York Shan and Lindsey Ran and Ming Hu and Valentina Emiliani and Na Ji and Jacob Reimer and Laurent Bourdieu and François St-Pierre},
howpublished = {HAL},
year = {2026},
month = jun,
doi = {10.64898/2026.06.01.726307},
url = {https://hal.science/hal-05827304v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - A red-emitting, genetically encoded indicator for two-photon voltage recording in vivo
AU - Shuyuan Yang
AU - Alex James Mcdonald
AU - Xiaoyu Lu
AU - Vincent Villette
AU - Noura Hakam
AU - Mario Galdamez
AU - Tanmayee Torne-Srivas
AU - Gregory Foran
AU - Xiaoyu Dong
AU - Matthew Shorey
AU - Nima Bavili
AU - Shujuan Lai
AU - Zhuohe Liu
AU - Haixin Liu
AU - Tom Maslianitsyna
AU - Rémi Fournel
AU - Emiliano Ronzitti
AU - Jun Zhu
AU - Ryan Gregory Natan
AU - Jonathan Bradley
AU - Daniela Gaspar Santos
AU - Yu-Yau York Shan
AU - Lindsey Ran
AU - Ming Hu
AU - Valentina Emiliani
AU - Na Ji
AU - Jacob Reimer
AU - Laurent Bourdieu
AU - François St-Pierre
PY - 2026
DA - 2026/06/03
PB - HAL
DO - 10.64898/2026.06.01.726307
UR - https://hal.science/hal-05827304v1
AB - Genetically encoded voltage indicators (GEVIs) enable minimally invasive, cell-type-specific optical measurements of neuronal membrane potential with millisecond temporal resolution. Red-shifted GEVIs are especially advantageous because they permit spectral multiplexing with complementary sensors and enable all-optical circuit interrogation in combination with blue-light-activated opsins. Despite these advantages, existing red GEVIs remain poorly suited for in vivo use due to limited performance under two-photon (2P) excitation, the predominant modality for deep-tissue imaging. Here, we introduce VADER1, a red GEVI that overcomes this limitation and enables reliable spike detection in vivo under 2P illumination. Under 2P excitation, VADER1 supports extended voltage imaging with both random-access and resonant-scanning microscopy, enables recordings from neurons as deep as cortical layer 5, and allows dual-color imaging with calcium indicators. By filling a critical spectral gap, VADER1 enables integrated optical measurements of fast electrical activity alongside other neural signals and establishes a foundation for two-photon all-optical electrophysiology.
ER -