The electrocardiogram (ECG) is the standard method in clinical practice to non-invasively analyze the electrical activity of the heart, from electrodes placed on the body’s surface. The ECG can provide a cardiologist with relevant information to assess the condition of the heart and the possible presence of cardiac pathology. Nonetheless, the global view of the heart’s electrical activity given by the ECG cannot provide fully detailed and localized information about abnormal electrical propagation patterns and corresponding substrates on the surface of the heart. Electrocardiographic imaging, also known as the inverse problem in electrocardiography, tries to overcome these limitations by non-invasively reconstructing the heart surface potentials, starting from the corresponding body surface potentials, and the geometry of the torso and the heart. This problem is ill-posed, and regularization techniques are needed to achieve a stable and accurate solution. The standard approach is to use zero-order Tikhonov regularization and the L-curve approach to choose the optimal value for the regularization parameter. However, different methods have been proposed for computing the optimal value of the regularization parameter. Moreover, regardless of the estimation method used, this may still lead to over-regularization or under-regularization. In order to gain a better understanding of the effects of the choice of regularization parameter value, in this study, we first focused on the regularization parameter itself, and investigated its influence on the accuracy of the reconstruction of heart surface potentials, by assessing the reconstruction accuracy with high-precision simultaneous heart and torso recordings from four dogs. For this, we analyzed a sufficiently large range of parameter values. Secondly, we evaluated the performance of five different methods for the estimation of the regularization parameter, also in view of the results of the first analysis. Thirdly, we investigated the effect of using a fixed value of the regularization parameter across all reconstructed beats. Accuracy was measured in terms of the quality of reconstruction of the heart surface potentials and estimation of the activation and recovery times, when compared with ground truth recordings from the experimental dog data. Results show that values of the regularization parameter in the range (0.01–0.03) provide the best accuracy, and that the three best-performing estimation methods (L-Curve, Zero-Crossing, and CRESO) give values in this range. Moreover, a fixed value of the regularization parameter could achieve very similar performance to the beat-specific parameter values calculated by the different estimation methods. These findings are relevant as they suggest that regularization parameter estimation methods may provide the accurate reconstruction of heart surface potentials only for specific ranges of regularization parameter values, and that using a fixed value of the regularization parameter may represent a valid alternative, especially when computational efficiency or consistency across time is required.