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. 2021 Dec 10;12:7229. doi: 10.1038/s41467-021-27444-9

Fig. 4. Two-photon imaging of optical field potential (OFP) with ArcLight variants in vivo.

Fig. 4

a Two-photon image of layer 2/3 neurons expressing ArcLight variants in V1. Average optical traces of entire field-of-view are shown in (b). Scale bar, 100 μm. b, c Fluorescence change of ArcLight variants (b) and stimulus-triggered average over 20 visual stimuli (c). Individual trials are shown with gray. Arrowheads indicate the timing of visual stimuli with flash light for 10 ms. d, e Peak ΔF/F (d) and SNR (e) of OFP in response to visual stimuli. Steel-Dwass test was used for statistical analysis. n = 40 events for each condition. Visual response was recorded 20 times in each FOV. Two mice were examined in each condition. f Two-photon image of primary visual cortex expressing Kv-ArcLight-ST under CAG promoter. Optical traces of small areas indicated with yellow dotted lines are shown in (g). Scale bar, 50 μm. g Fluorescence change of Kv-ArcLight-ST. h Magnified traces in the area shown with a red rectangle in (g). Optical signals of neighboring areas showed similar dynamics. i Cross-correlation between areas. j Relationship between correlation and distance areas. Two-photon imaging was performed in anaesthetized mice with isoflurane (1.5% v/v). Excitation: 940 nm, 130–150 mW (0.50–0.61 μW/pixel) at the imaging plane. In each box plot, the central line represents the median, and the bottom and the top edges of the box represent the 25th and 75th percentile, respectively. The lower and the upper whiskers extend to the minimum and the maximum data points, respectively. Numbers on the graphs represent P values.