Automatic quantification of regional left ventricular deformation in volumetric ultrasound data remains challenging. Many methods have been proposed to extract myocardial motion, including techniques using block matching, phase-based correlation, differential optical flow methods, and image registration. Our lab previously presented an approach based on elastic registration of subsequent volumes using a B-spline representation of the underlying transformation field. Encouraging results were obtained for the assessment of global left ventricular function, but a thorough validation on a regional level was still lacking. For this purpose, univentricular thick-walled cardiac phantoms were deformed in an experimental setup to locally assess strain accuracy against sonomicrometry as a reference method and to assess whether regions containing stiff inclusions could be detected. Our method showed good correlations against sonomicrometry: r 2 was 0.96, 0.92, and 0.84 for the radial (ε RR ), longitudinal (ε ), and circumferential (ε CC LL ) strain, respectively. Absolute strain errors and strain drift were low for ε (absolute mean error: 2.42%, drift: −1.05%) and ε CC LL (error: 1.79%, drift: −1.33%) and slightly higher for ε (error: 3.37%, drift: 3.05%). The discriminative power of our methodology was adequate to resolve full transmural inclusions down to 17 mm in diameter, although the inclusion-to-surrounding tissue stiffness ratio was required to be at least 5:2 (absolute difference of 39.42 kPa). When the inclusion-to-surrounding tissue stiffness ratio was lowered to approximately 2:1 (absolute difference of 22.63 kPa), only larger inclusions down to 27 mm in diameter could still be identified. Radial strain was found not to be reliable in identifying dysfunctional regions.