Fig. 6: JEDI-1P-Kv reports rapid voltage responses to high-frequency stimuli.

a–c Simultaneous single whisker deflection and widefield voltage imaging in awake mice. a Experimental schematics. b Contralateral barrel cortex shows rapid response to a single whisker deflection. n = 150 deflections from an animal, representative of results with 5 mice. Responses of the other 4 mice are shown in Supplementary Fig. 11. c Example time series shows rapid responses to single whisker deflections in a single trial. n = 1 mouse, representative of results with 5 mice. d–g Simultaneous visual stimulation and widefield voltage imaging in awake mice. d Experimental schematics. e 40-Hz-filtered (or 60-Hz) optical responses to 40-Hz (or 60-Hz) visual stimulations are bilaterally localized in the visual cortex. The heat map shows the mean power difference at 40 Hz (or 60 Hz) between stimulation and baseline. n = 4 mice, 10 trials/mouse. f Single-trial imaging of JEDI-1P-Kv can report local voltage responses at frequencies matching visual stimulations of up to 60 Hz. Responses were acquired from a 2 × 2 pixel area in the visual cortex (arrow in (e)). n = 1 mouse, representative of results with 4 mice. g Oscillations in the responses to specific flicker frequencies are visible in averaged voltage signals up to 60 Hz. Data from a 2 × 2 pixel area in the primary visual cortex. Data shown were acquired 1.5–2.0 s into a 3-s stimulus train. n = 4 mice, 10 trials/mouse. h–k Simultaneous whisker stimulation and widefield voltage imaging in awake mice. h Experimental schematic. Air-puff trains were applied to the right set of whiskers. i 40-Hz-filtered (or 60-Hz) optical responses to 40-Hz (or 60-Hz) air puffs are localized at the contralateral barrel cortex. The heat map shows the mean power difference at 40 Hz (or 60 Hz) between stimulation and baseline. n = 5 mice, 10 trials/mouse. j Single-trial imaging of JEDI-1P-Kv can report local voltage responses at frequencies matching air-puff stimulations of up to 60 Hz. Responses were acquired from a 2 × 2-pixel area in the contralateral barrel cortex (arrow in (i)). n = 1 mouse, representative of results with 5 mice. k Same as (g) but for barrel cortex responses to somatosensory stimuli. Data shown were acquired 1.5–2.0 s into a 3-s stimulus train. n = 5 mice, 10 trials/mouse. l, m Responses to air puffs are stable over multiple days and imaging sessions. Neural responses are delayed compared with the valve opening triggers (dashed lines) due to the ~7-ms valve opening time. n = 216 trials from 6 mice (36 trials/mouse). Gray traces were acquired 16–19 imaging sessions after the orange and purple traces. The initial larger peak is consistent with electrophysiological data⁹³. Dark traces are the mean, and shaded areas denote the 95% CI. Panels (a, d, and h) were created using Biorender.com.