FIGURE 3

pyHeart4Fish validation and interpretation guide (A) Pearson correlation analysis for manual and semi-automated heart rate measurements in atrium and ventricle using pyHeart4Fish (pyH4F) vs. manual determination (50 zebrafish), user-dependent determination (85 zebrafish), and pairwise comparison of replicates (561 zebrafish, 2–13 replications). Outliers (shown in gray) with more than 3-fold y standard deviation from the regression curve were not considered when calculating the Pearson correlation coefficient. (B) An interpretation guide for max. dilation and max. contraction data obtained with pyHeart4Fish. The smallest circles indicate max. contraction and circles larger than these indicate dilation. Blue represents controls and orange represents treated hearts. The arrows indicate the difference between min. and max. chamber size in treated fish. The displayed examples demonstrate increased max. dilation with decreased max. contraction after opipramol diHCl exposure, and reduced max. dilation and contraction in the atrium of benzydamine HCl-treated embryos (C) A representative raw data output of pyHeart4Fish analysis. The overlay of raw-signal in combination with the arrhythmia score, and conduction score shows there are no conduction defects between the atrium and ventricle in opipramol diHCl- and benzydamine HCl-treated embryos. Black line = raw signal; red dotted line = fitted sine function; freq = frequency estimated using the sine function; fft_freq = fast fourier transformation frequency; HCl = hydrochloride.