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June 6th, 2025
Version: 2
Peking University
bioengineering
biorxiv

A photostable genetically encoded voltage indicator for imaging neural activities in tissue and live animals

Cao, C.Open in Google Scholar•Zhu, R.Open in Google Scholar•Zhou, S.Open in Google Scholar•Zhao, Z.Open in Google Scholar•Lin, C.Open in Google Scholar•Liu, S.Open in Google Scholar•Peng, L.Open in Google Scholar•Subach, F. V.Open in Google Scholar•Piatkevich, K. D.Open in Google Scholar•Zou, P.Open in Google Scholar

Genetically encoded voltage indicators (GEVIs) enable noninvasive, highspeed monitoring of electrical activity but are constrained by limited brightness and rapid photobleaching under continuous illumination. Here, we present Vega, a highly photostable green fluorescence GEVI with both high sensitivity ({Delta}F/F{square}={square}-33% per 100{square}mV) and fast response (1.34{square}ms). Under onephoton excitation at 1 W/cm2, Vega exhibits more than 20-fold slower photobleaching than the spectrally similar GEVI, Ace-mNeon2. In acute mouse brain slice, Vega enabled widefield high-fidelity recording of action potentials from 51 neurons simultaneously. In pancreatic islets, it revealed heterogeneous {beta}-cell activation and intercellular coupling in response to glucose elevation. Finally, onephoton imaging in awake mice demonstrated stable cortical voltage mapping in vivo. Vega thus overcomes the longstanding photostability-performance tradeoff, enabling chronic, highfidelity voltage imaging across preparations.

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