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A versatile, positive-going voltage indicator that enables accessible two-photon recordings in vivo | bioRxiv

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Genetically encoded voltage indicators (GEVIs) enable cell-type–specific optical readout of membrane potential, but two-photon (2P) spike detection has been hampered by low signal-to-noise and ultrafast off-kinetics, restricting use to specialized microscopes. We introduce FORCE1s, a green, positive-going GEVI engineered to make robust 2P voltage imaging broadly accessible. FORCE1s brightens from a dark baseline during depolarization, reports spikes with ∼100% ΔF/F in awake mice, and displays repolarization kinetics that are tuned for reliable spike detection at sub-kilohertz frame rates. As a result, FORCE1s supports spike-resolved multi-cell recordings on standard resonant-scanning microscopes, and further scales to larger fields of view and neuron counts on advanced modalities. FORCE1s also enables multiplexed voltage–neurotransmitter imaging and extended recordings in freely moving mice using a compact, affordable MEMS-based 2P miniscope. Together, these advances establish FORCE1s

New Results , Michelle Ann Land, Shuyuan Yang, Noura Hakam, Vincent Villette, Jun Zhu, Mario Galdamez, Mario Fernandez de La Puebla, Xiaoyu Lu, Gregory Foran, Tanmayee Torne-Srivastava, Beatriz Campillo, Haixin Liu, Xiaoyu Dong, Shujuan Lai, Matthew Shorey, Hala Abdallah, Raegan Banks, Anastasia Mamontova, Yu-Yau York Shan, Ryan Kroeger, Robert G. Law, Ming Hu, Daniela Gaspar Santos, Jonathan Bradley, Alberto Lombardini, Benjamin Mathieu, Annick Ayon, Ryan Gregory Natan, Hongyi Yuan, Jacob Reimer, Laurent Bourdieu, Na Ji, Weijian Zong, View ORCID ProfileFrançois St-Pierre doi:…

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