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63) than SE (2.56, p = 0.01), and state-of-the-art 2-D digital image correlation (1.81, p less then 0.001) and direct image subtraction (1.29, p less then 0.001) methods. Histopathology agreed well with ultrasound (Dice coefficient = 0.48), with deviations mainly explained by the three-dimensional inhomogeneous distribution of BTX-A. Preliminary in vivo patient results indicated that SF and SE discard artifactual BTX-A detection outside the injection region. The proposed methods contribute to objectivize ultrasound-guided injections, w