GEVIBench › All sensors › rEstus
rEstus
Variant of ASAP3 optimized for non-excitable cells, shifting voltage sensitivity to match their characteristically negative resting potentials.
- Category: VSD-cpGFP GEVI
- Published: 2024-03-25 — Advanced Science 2024
- Derived from: ASAP3
- Response polarity: negative-going (fluorescence decreases on depolarization)
- Spectrum: excitation peak 463 nm, emission peak 513 nm (rEstus)
- Plasmid: Addgene #209767
- Tags: cpGFP
Headline values (as shown in the GEVIBench table)
| Metric | Value | Definition |
|---|---|---|
| Brightness (B/B_EGFP) | 0.85× EGFP | Molecular brightness relative to EGFP (extinction coefficient × quantum yield, or a measured ratio). Sensors not measured against EGFP directly are placed through a chain of pairwise brightness comparisons, combined by geometric mean. |
| Kinetics τ_on / τ_off (ms) | 14 / 51 ms | Activation / decay time constants in ms (amplitude-weighted mean τ for multi-exponential fits). The value shown averages the measurements made at physiological temperature (≥33 °C); if there are none, it is the measurement closest to 33 °C. |
| ΔF/F per 100 mV | -62% | Steady-state fluorescence change for a 100 mV depolarization step (−70 → +30 mV); the sign gives the polarity. The table shows the first listed measurement; the median across measurements (one-photon preferred) is used for sorting. |
| ΔF/F per mV | 1.3% | Subthreshold sensitivity: fluorescence change per mV near rest (−90 to −50 mV). Where no value is reported directly, it is estimated from the F–V curve. |
| ΔF/F per AP | — | Fluorescence change per single action potential in neurons. The table shows the first listed measurement; the median across measurements (one-photon preferred) is used for sorting. |
| AP width (FWHM, ms) | — | Full width at half maximum of the optical spike waveform. |
| Photostability t75% @ 100 mW/mm² | — | Time for fluorescence to fall to 75% of its initial value under one-photon illumination, linearly dose-scaled to 100 mW/mm² (t50% is used where the 75% crossing falls inside a fast initial transient). Two-photon curves are not dose-scaled and are excluded. |
| Papers using it (N_used) | 5 | Number of published research papers, tracked by GEVIBench, that used the sensor for an experiment (a popularity / track-record measure). |
Measurements and sources
Kinetics
- τ_on 13.5 ms, τ_off 51 ms at 23°C — Rühl 2024, Fig. S9
ΔF/F per 100 mV
- -62%, 1P — Rühl 2024, Fig. 1G
ΔF/F per mV (subthreshold)
- 1.25%/mV (Peak fractional voltage sensitivity (dF/F dV-1, %/mV) read from Fig. S2 of the rEstus2s paper, the first derivative of the Boltzmann F-V fit divided by the fit; rEstus (black) peaks at ~1.25 %/mV near -10 to 0 mV. Re-sourced here because the origin paper's Fig. 1H plots absolute |dB/dVm| in %EGFP/mV (a different quantity), not fractional %/mV. On the same axis the sibling rEstus2s (red) peaks at ~2.0 %/mV near +5 mV (about 1.6-fold over rEstus across the -100 to 0 mV resting range).) — Rühl 2026, Fig. S2
Brightness comparisons
- 0.775× the brightness of EGFP — Rühl 2024, Fig. 1G
- 2.07× the brightness of ASAP3 — Rühl 2024, Table 1
- 1.71× the brightness of ASAP3 — Nair 2025, Table S1
- 0.9569377990430623× the brightness of rEstus-NI — Nair 2025, Fig. 2C
- 1.669449081803005× the brightness of rEstus2s — Rühl 2026, Table S1
Fluorescence–voltage (F–V) curve
ΔF/F (%) at each membrane potential, normalized to 0 at −70 mV: -120 mV: 13%; -100 mV: 10%; -80 mV: 5%; -70 mV: 0%; -60 mV: -6%; -40 mV: -20%; -20 mV: -36%; 0 mV: -50%; 20 mV: -59%; 40 mV: -64%; 60 mV: -67%; 75 mV: -68% — Rühl 2024, Fig. 1G
Research papers using rEstus (5)
- An Ultrasensitive Genetically Encoded Voltage Indicator Uncovers the Electrical Activity of Non-Excitable Cells — Rühl P, et al., Advanced Science 2024 (HEK293T cells; A375 melanoma cells; MCF-7 breast cancer cells, in vitro)
- Sub-Millivolt Voltage Imaging Reveals Gap Junction-Mediated Bioelectric Contact Inhibition — Rühl P, et al., bioRxiv 2026 (HEK293T cells, in vitro (rEstus characterized under whole-cell voltage clamp alongside rEstus2s))
- Absolute Membrane Potential Recording with ASAP-Type Genetically Encoded Voltage Indicators Using Fluorescence Lifetime Imaging — Nair AG, et al., ACS Chemical Neuroscience 2025 (HEK293T cells; A375 melanoma cells; MCF-7 breast cancer cells, in vitro)
- Crosstalk of KCNH1 and KCNH5 gain-of-function mutations leading to epilepsy and neurodevelopmental disorders — Bernert A, et al., Molecular Brain 2026 (HEK293T cells, in vitro)
- Generation and characterization of a human induced pluripotent stem cell line (hiPSC) expressing the rEstus voltage sensor under doxycycline induction — Malan D, et al., Stem Cell Research 2026 (Human iPSC, in vitro)
Data and citation
- llms.txt — summary for language models
- llms-full.txt — every sensor in full (Markdown)
- gevis.json — all values and measurements with sources (JSON)
- gevis.csv — one row per sensor (CSV)
Cite GEVIBench as: Adam E. Cohen and Daozheng Gong (2026). Voltage imaging: A practical guide. Neuron. https://doi.org/10.1016/j.neuron.2026.08.002
Generated from the GEVIBench database on 2026-10-12.