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Background,Motivation and Objective Fiber architecture is of major interest in the cardiac function.Recently,Backscatter Tensor Imaging(BTI),a new imaging technique enabled the detection of fiber organization in ex vivo myocardial samples using spatial coherence measurement on a linear array.In this study,3D BTI was developed on a 2D matrix probe to calculate 2D coherence function(CF) to retrieve the 3D fiber architecture dynamic in vivo on a beating heart of an open-chest sheep during an entire cardiac cycle.Moreover,we validated the fiber orientations derived with 3D BTI against histology.