Mapping 3D fiber orientation in tissue using dual-angle optical polarization tractography.

Optical polarization tractography (OPT) has recently been applied to map fiber organization in the heart, skeletal muscle, and arterial vessel wall with high resolution. The fiber orientation measured in OPT represents the 2D projected fiber angle in a plane that is perpendicular to the incident light. We report here a dual-angle extension of the OPT technology to measure the actual 3D fiber orientation in tissue. This method was first verified by imaging the murine extensor digitorum muscle placed at various known orientations in space. The accuracy of the method was further studied by analyzing the 3D fiber orientation of the mouse tibialis anterior muscle. Finally we showed that dual-angle OPT successfully revealed the unique 3D "arcade" fiber structure in the bovine articular cartilage.