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Figure 3 Comparison of heart rate and cardiac rhythm measurement in control zebrafish embryos aged at 72 hpf using either OpenCV or ImageJ methods. (A) Schematic diagram showing the basic definition for atrium and ventricle signals. (B) Comparison of heart rate obtained from either atrium (left panel) or ventricle (right panel) chambers using either ImageJ or OpenCV methods. (C) Comparison of the atrium to atrium (A-A) and ventricle to ventricle (V-V) intervals using either ImageJ or OpenCV methods. (D) Comparison of atrium-ventricle (A-V) and ventricle to atrium (V-A) intervals using either ImageJ or OpenCV methods. Comparison of atrium-atrium (A-A) (E) and ventricle-ventricle (V-V) (F) interval regularity using either ImageJ or OpenCV methods. Data was processed using unpaired t-test (B–D) as it follows the normal distribution. Pearson correlation (B) and Wilcoxon test (E,F) were conducted because the data does not obey normal distribution (*** p < 0.001, **** p < 0.0001).