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. 2018 Feb 22;8:3493. doi: 10.1038/s41598-018-21640-2

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© The Author(s) 2018

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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FISSA performance for different simulated data parameters and user-adjustable FISSA parameters, for the signal of the cell of interest from the case in Fig. 2C. (A) Correlations between the source and extracted signals before neuropil decontamination (‘Measured’) and after each decontamination method (neuropil subtraction, cNMF, and FISSA) for different simulation parameters. (i) Correlations for changes in the firing rate of the cell of interest (0.1 to 1.7 Hz, with steps of 0.2 Hz, default is 0.3 Hz). (ii) Correlations for changes in calcium transient magnitude (parameter A, see Eq. 9. 0.06 to 0.54 with steps of 0.06, default is 0.3). (iii) Correlations for changes in imaging framerate (downsampling of 100 Hz initial data to lower frame rates: by 50, 40, 30, 20, 10, 5, 4, 3, 2, and 1). (iv) Correlations for changes in cell shape, by changing the parameter ρ, see Eq. 10 (steps of 5 from 0 to 45, default is 0). The insets show example cell shapes for different ρ values. (B) Correlations between the source and extracted signals against different user-adjustable FISSA parameters (x-axes). (i) Number of neuropil subregions while keeping the total area constant, at four times the ROI area. (ii) Area of the neuropil subregions relative to the central ROI (0.025 for the smallest area, steps of 0.5 from 0.5 to 4), for four subregions. (iii) NMF parameter α (with steps of 0.1). (C) The effect of suboptimal ROI selection, by varying the threshold at which a cell mask is defined (Parameter $T_{mask}$ in Eq. 14. 0.1 to 1.1 with steps of 0.1, default is 0.5). The insets show the same central cell with the outlines of the mask used for three example thresholds. All panels show the average values over 10 simulations; shaded areas indicate standard error.