ArticleScience advances2025
Photophysics-informed two-photon voltage imaging using FRET-opsin voltage indicators.
Article in Science advances, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- Crosstalk-free optical indicator-actuator combinations for bidirectional optogenetics.npj biomedical innovations · 2026Review
- Absolute voltage mapping using dynamic photocycle control.bioRxiv : the preprint server for biology · 2026Article
- Improving positively tuned voltage indicators for faster kinetics and higher contrast.bioRxiv : the preprint server for biology · 2026Article
- Noise Sources and Strategies for Signal Quality Improvement in Biological Imaging: A Review Focused on Calcium and Cell Membrane Voltage Imaging.Biosensors · 2026Review
- Miniaturized widefield microscope for high speed in vivo voltage imaging.Biomedical optics express · 2026Article
- Voltage imaging with periodic structured illumination.Biomedical optics express · 2026Article
- Miniaturized widefield microscope for high speed in vivo voltage imaging.bioRxiv : the preprint server for biology · 2025Article
- Voltage Imaging with Periodic Structured Illumination.bioRxiv : the preprint server for biology · 2025Article
- Robotic Fast Patch Clamp in Brain Slices Based on Stepwise Micropipette Navigation and Gigaseal Formation Control.Sensors (Basel, Switzerland) · 2025Article
- A novel rhodopsin-based voltage indicator for simultaneous two-photon optical recording with GCaMP in vivo.bioRxiv : the preprint server for biology · 2024Article
- Optical constraints on two-photon voltage imaging.Neurophotonics · 2024Article
- Optical constraints on two-photon voltage imaging.bioRxiv : the preprint server for biology · 2024Article
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6 authors.
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Abstract
Microbial rhodopsin-derived genetically encoded voltage indicators (GEVIs) are powerful tools for mapping bioelectrical dynamics in cell culture and in live animals. Förster resonance energy transfer (FRET)-opsin GEVIs use voltage-dependent quenching of an attached fluorophore, achieving high brightness, speed, and voltage sensitivity. However, the voltage sensitivity of most FRET-opsin GEVIs has been reported to decrease or vanish under two-photon (2P) excitation. Here, we investigated the photophysics of the FRET-opsin GEVIs Voltron1 and Voltron2. We found that the previously reported negative-going voltage sensitivities of both GEVIs came from photocycle intermediates, not from the opsin ground states. The voltage sensitivities of both GEVIs were nonlinear functions of illumination intensity; for Voltron1, the sensitivity reversed the sign under low-intensity illumination. Using photocycle-optimized 2P illumination protocols, we demonstrate 2P voltage imaging with Voltron2 in the barrel cortex of a live mouse. These results open the door to high-speed 2P voltage imaging of FRET-opsin GEVIs in vivo.
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