rEstus-NI

Variant of rEstus optimized for fluorescence lifetime imaging and absolute membrane potential measurement.

Category: VSD-cpGFP GEVI
Published: 2025-12-17 — ACS Chemical Neuroscience 2025
Derived from: rEstus
Response polarity: negative-going (fluorescence decreases on depolarization)
Spectrum: excitation peak 463 nm, emission peak 513 nm (rEstus-NI)
Tags: cpGFP

Headline values (as shown in the GEVIBench table)

Metric	Value	Definition
Brightness (B/B_EGFP)	0.84× 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)	181 / 433 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	-57%	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	1.1% (estimated from the F–V curve)	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)	1	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 181 ms, τ_off 433 ms at 20-23°C — Nair 2025, Fig. S6C

ΔF/F per 100 mV

-57%, 1P — Nair 2025, Fig. 1F

Brightness comparisons

1.045× the brightness of rEstus — Nair 2025, Fig. 2C
1.788× the brightness of ASAP3 — Nair 2025, Fig. 2C

Fluorescence–voltage (F–V) curve

ΔF/F (%) at each membrane potential, normalized to 0 at −70 mV: -120 mV: 66%; -100 mV: 40%; -80 mV: 11%; -70 mV: 0%; -60 mV: -11%; -40 mV: -30%; -20 mV: -42%; 0 mV: -51%; 20 mV: -55%; 40 mV: -58%; 60 mV: -61% — Nair 2025, Fig. 1F

Research papers using rEstus-NI (1)

Absolute Membrane Potential Recording with ASAP-Type Genetically Encoded Voltage Indicators Using Fluorescence Lifetime Imaging — Nair AG, Rodewald M, Bae H, Rühl P, Popp J, Schmitt M, Meyer-Zedler T, Heinemann SH, ACS Chemical Neuroscience 2025 (HEK293T cells; A375 melanoma cells; MCF-7 breast cancer cells, 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.