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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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Onset, duration and decay—calcium (GECI) versus voltage (GEVI) optical signals. (A1) Transgenic animal expressing GECI (GCaMP6f) in cortical pyramidal neurons. Coronal brain slice imaged by NeuroCCD 80 × 80 pixel. Scale, 200 µm. Two ROIs are selected, at the stimulation site (ROI-1), and 200 µm away from the stimulation site (ROI-2). (A2) Synaptically-evoked optical signals in GECI mouse. Time-to-peak parameter is measured between stimulus pulse and calcium signal peak. Signal duration is measured in the 3rd peak at half amplitude (Half-Width). (B1) Same as (A1), except the transgenic animal is expressing GEVI (chi-VSFP). (B2) Same as in (A2), except GEVI mouse. (C) Each dot represents one measurement (one experimental trial) of the time-to-peak parameter quantified at the stimulation site (ROI-1). Calcium: 44 trials, in 6 brain slices from 3 animals. Voltage: 60 trials, in 13 slices, of 7 animals. ***, p < 0.0001. (D) Same as in (C), except different parameter, half-width. (E1) Calcium (black) and voltage (green) optical signals are superimposed on the same time scale. Ca signal decreases 50% in time interval marked by double arrow. Vertical turquois line marks the time point “144.5 ± 10.5 ms”, when, on average, Ca signal amplitudes dropped down to 50% of their maxima. At that moment of time, “144.5 ± 10.5 ms”, Ca signal is at 50%, while the voltage trace is near the baseline (0–5%). (E2) Same as in (E1), except different ROI (away from the stimulation site, ROI-2). Voltage signal decreases by 90% in time interval marked by double arrow. (F) Three optical signals from 3 GECI animals, followed by 3 optical signals from 3 GEVI animals. Optical signals are aligned by synaptic stimulation pulses (syn.). Bottom: Green data points: time intervals (from the synaptic stimulus) at which voltage signals dropped down to 10% of their maxima (V max). The average time delay for this 90% descent was 81 ± 7 ms (n = 29). Vertical green arrow marks the time point “81 ± 7 ms” transecting voltage and calcium waveforms. Bottom, orange points: Ca signal amplitudes measured 81 ms after the syn. pulse; normalized to the “Ca max” of the same trace; 85 ± 6% (n = 54). The voltage had dropped down to 10%, while the calcium is still lingering at ~ 85%.