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JEDI-2P

Two-photon optimized GEVI evolved from ASAP2s with improved brightness, photostability, and sensitivity under two-photon illumination.

Category: VSD-cpGFP GEVI
Published: 2022-08-18 — Cell 2022
Derived from: ASAP2s
Response polarity: negative-going (fluorescence decreases on depolarization)
Spectrum: excitation peak 501 nm, emission peak 519 nm (JEDI-2P)
Two-photon compatible: yes
Plasmid: Addgene #179463
Tags: cpGFP, Two-photon

Headline values (as shown in the GEVIBench table)

Metric	Value	Definition
Brightness (B/B_EGFP)	0.51× 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)	2.1 / 2.8 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	-48% (median of 5 measurements: −45.5%)	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	0.7%	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	18.2% (median of 6 measurements: 24%)	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)	1.6 ms	Full width at half maximum of the optical spike waveform.
Photostability t75% @ 100 mW/mm²	14 s	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)	14	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 1.45 ms, τ_off 1.21 ms at 32-35°C — Liu 2022, Table S2
τ_on 2.47 ms, τ_off 2.54 ms at 21-23°C — Liu 2022, Table S2
τ_on 0.91 ms, τ_off 1.5 ms at 33°C — McDonald 2026, Table S1.4
τ_on 1.6 ms, τ_off 1.2 ms at 33°C — Land 2026, Table 1
τ_on 4.57 ms, τ_off 3.8 ms at 23°C — Hao 2024, Table S2
τ_on 4.3 ms, τ_off 7.28 ms at 37°C — Hao 2024, Table S2

AP width (FWHM)

1.4 ms at 7142.9 Hz sampling, Mouse, visual cortex L2/3, awake head-fixed, in vivo, 2P, 37°C — Yang 2026, Fig. 3C
5.6 ms at 20000 Hz sampling, HEK293A cells, whole-cell voltage clamp, in vitro (2-ms FWHM command AP waveform), 2P, 23°C — Land 2026, Fig. 1k
1.4 ms at 1000 Hz sampling, Mouse visual cortex, awake behaving, in vivo, soma-targeted JEDI-2P-Kv (ULoVE 2P), 2P, 37°C — Land 2026, Fig. 2d
1.4 ms at 7100 Hz sampling, Mouse, visual cortex L2/3, awake head-fixed, in vivo (ULoVE 2P), soma-targeted JEDI-2P-Kv, 2P, 37°C — McDonald 2026, Fig. 3E
3.6 ms at 721 Hz sampling, HEK293A cells, whole-cell voltage clamp, in vitro (2-ms FWHM command AP waveform), 2P, 33°C — McDonald 2026, Fig. 1K
2.1 ms at 4400 Hz sampling, Mouse, visual cortex L2/3, anesthetized, in vivo (2P, paired juxtacellular), 2P, 37°C — Liu 2022, Fig. 5D
1.84 ms at 3500 Hz sampling, Mouse, V1 cortex layer II/III, awake behaving, in vivo (JEDI-2P-Kv, ULoVE 2P), 2P, 37°C — Villette 2024, Fig. 3f
0.72 ms at 3500 Hz sampling, Mouse, cerebellar granular layer, fast-spiking Glyt2+ Golgi interneurons, awake behaving, in vivo (JEDI-2P, ULoVE 2P), 2P, 37°C — Villette 2024, Fig. S9

Brightness comparisons

1.67× the brightness of ASAP3 — Liu 2022, Fig. 2I
1.5× the brightness of ASAP2s — Liu 2022, Fig. 2I
1.45× the brightness of ASAP5 — Land 2026, Ext. Fig. 1c
0.24271844660194175× the brightness of 2Photron — Villette 2024, Fig. 3f
2.564102564102564× the brightness of FORCE1s — McDonald 2026, Fig. 1C
0.24509803921568626× the brightness of Jarvis — Grimm 2026, Fig. 1C
1.1627906976744187× the brightness of JEDI-3hyp — Land 2026, Fig. 1d
1.0416666666666667× the brightness of JEDI-3sub — Land 2026, Fig. 1d

Photobleaching

t75% = 64 s at 37 mW at sample plane (2P), 2P — Land 2026, Fig. 1e
t75% = 44 s, 1P — Grimm 2026, Fig. 1F
t75% = 14 s at 100 mW at sample plane (2P, 940 nm), 2P — McDonald 2026, Fig. S1.2B
t75% = 234 s at 22.5 mW at 1045 nm (ULoVE 2P, brain surface), 2P — Yang 2026, Fig. 3F
t50% = 65 s, 1P — Liu 2022, Fig. S4G

Fluorescence–voltage (F–V) curve

ΔF/F (%) at each membrane potential, normalized to 0 at −70 mV: -100 mV: 10%; -80 mV: 3%; -60 mV: -5%; -40 mV: -20%; -20 mV: -34%; 0 mV: -43%; 20 mV: -47%; 30 mV: -48%; 50 mV: -49% — Liu 2022, Fig. 2B

Research papers using JEDI-2P (14)

Sustained deep-tissue voltage recording using a fast indicator evolved for two-photon microscopy — Liu Z, Lu X, Villette V, et al., Cell 2022 (HEK293 cells, in vitro; Mouse, visual cortex L2/3, in vivo; Drosophila, interneurons, ex vivo; Mouse, retina, ex vivo)
Scanless two-photon voltage imaging — Sims RR, et al., Nature Communications 2024 (Mouse, hippocampal slices, in vitro; Mouse, barrel cortex, in vivo)
A fast and responsive voltage indicator with enhanced sensitivity for unitary synaptic events — Hao YA, et al., Neuron 2024 (Rat, cultured neurons, in vitro; Mouse, cortex, in vivo)
A novel rhodopsin-based voltage indicator for simultaneous two-photon optical recording with GCaMP in vivo — Villette V, et al., bioRxiv 2024 (Mouse, visual cortex, in vivo)
Two-photon voltage imaging with rhodopsin-based sensors — Grimm C, et al., Neuron 2026 (HEK293 cells, in vitro; Mouse, cortex and cerebellum, in vivo)
All-optical voltage interrogation for probing synaptic plasticity in vivo — Carolan J, et al., Nature Communications 2025 (Mouse, cerebellum, Purkinje cell dendrites, in vivo)
FACED 2.0 enables large-scale voltage and calcium imaging in vivo — Zhong J, et al., Nature Methods 2025 (Mouse, visual cortex L2/3, in vivo; Zebrafish larvae, in vivo)
Retroreflective beam multiplexing enables high-throughput two-photon voltage imaging in vivo — Liu Z, et al., bioRxiv 2025 (Mouse, visual cortex V1, in vivo; Mouse, hippocampus CA1, in vivo)
Movement-stabilized three-dimensional optical recordings of membrane potential changes and calcium dynamics in hippocampal CA1 dendrites — Gonzalez KC, et al., Neuron 2026 (Mouse, hippocampus CA1, dendrites and soma, in vivo)
Kilohertz-rate two-photon voltage imaging of population dynamics in vivo — Zhang M, et al., bioRxiv 2026 (Mouse, cortex, in vivo)
Designer indicators for two-photon recording of subthreshold voltage dynamics — Land MA, Galdamez M, Villette V, Zhu J, Lu X, Marosi M, et al., Nature Methods 2026 (HEK293 cells, in vitro; Rat, cultured hippocampal neurons, in vitro; Mouse, visual cortex L2/3, in vivo)
Optical constraints on two-photon voltage imaging — Brooks FP, et al., Neurophotonics 2024 (HEK293 cells, in vitro; Mouse, cortex, in vivo)
Linking functional activity and connectivity of neuronal circuits via fast cross-layer all-optical physiology — Liu C, et al., Cell Reports 2025 (Mouse, visual cortex V1 and somatosensory cortex S1, in vivo)
A red-emitting, genetically encoded indicator for two-photon voltage recording in vivo — Yang S, McDonald AJ, Lu X, Villette V, Hakam N, Galdamez M, et al., bioRxiv 2026 (Mouse, visual cortex L2/3, in vivo (JEDI-2P-Kv used as the in vivo benchmark for the red GEVI VADER1 under ULoVE and resonant-scanning 2P microscopy))

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.