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. 2021 Mar 5;11:5295. doi: 10.1038/s41598-021-84651-6

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Optical signal propagation—calcium vs. voltage imaging. (A) Top, image: Surface of a cortical brain slice prepared from GCaMP6f animal, with ROIs and glass electrode for synaptic stimulation. Bottom, traces: Synaptically-evoked calcium (GECI) transients in the presence of GABA-A receptor antagonist, gabazine, GBZN [10 µM]. Synaptic stimulation comprised two triplets, 8.3 Hz (Train-1) and 83 Hz (Train-2), respectively. Optical traces were recorded simultaneously from 5 ROIs. The remote traces (ROI-3-4-5) are shown at two amplifications: × 1 (black) and × 8, × 16 (color). Asterisk marks uneventful waveform. (B) Same as in (A), except the brain slice was prepared from a chi-VSFP animal. Black arrows mark fast changes in GEVI waveforms, which are unmatched in the GECI records (asterisk). Inset: Differences in the peak timing and signal duration between a GECI and GEVI trace at ROI-2. (C) Traces from (A) and (B) (Train-2) are amplitude-scaled, time-aligned, and superimposed on a faster time scale. Yellow box on the rising slopes of traces indicates the amplitude level (half-amplitude) at which latencies were quantified. Note that voltage transients (bottom) show a greater variety of signal latencies compared to the calcium transients (top). Vertical dashed line “onset” marks the onset of synaptic stimulation. Vertical dashed line “V-peak” marks the peak of the most delayed voltage transient (ROI-3). Thick vertical line “Ca-peak” marks the peak of the GCaMP6f transients. (D) Inset: Propagation latency between two ROIs was measured at half-amplitude. White bar is an average latency measured at 500 µm away from the stimulation site, obtained in 21 recordings from 6 brain slices treated with gabazine, in 3 GCaMP animals (mean ± stdev). Gray bar is an average 500 µm latency obtained in 37 recordings from 15 GBZN-treated brain slices, in 6 chi-VSFP animals, p < 0.00001.