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. 2019 Jan 15;9:145. doi: 10.1038/s41598-018-35949-5

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© The Author(s) 2019

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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A comparison of heart rates derived from automated placement of Region of Interest match those derived via manual placement. (a) Heart rate data from both ROI placement methods were plotted against one another and linear regression derived r², slope and p-value. r value indicates correlation of manual and automatic placement of ROI-derived data of all imaged animals across 10 total experiments. The line of identity (blue) illustrates a hypothetically “perfect” correlation between the two data sets for comparison. (b) A Bland-Altman analysis provides quantitative data on the disagreement between the two methods (bias) and the interval within which 95% of all disagreement occurs (95% limits of agreement indicated by dashed lines; −5.346 to 5.594). The x-axis is the average between the heart rates derived from the two methods, and the y-axis is the distance that those two values are from one another (Method A-Method B). A positive value on the y-axis indicates that Method A (manual ROI placement) returned a higher value, and vice versa.