To determine the relation between local myofiber anatomy and local deformation in the wall of the left ventricle, both three-dimensional transmural deformation and myofiber orientation were examined in the anterior free wall of seven canine left ventricles. Deformation was measured by imaging columns of implanted radiopaque markers with high-speed, biplane cineradiography (16 mm, 120 frames/sec). Hearts were fixed at end diastole and sectioned parallel to the local epicardial tangent plane to determine the transmural distribution of fiber directions at the site of strain measurement. The principal direction of deformation associated with the greatest shortening was compared with the local fiber direction in the outer (21 +/- 8% of the wall thickness from the epicardium) and inner (65 +/- 9%) halves of the wall. Although the fiber direction varied substantially with depth from the epicardium, the principal direction did not. In the outer half of the wall, fiber direction averaged -8 +/- 24 degrees, while the principal direction averaged -33 +/- 24 degrees from circumferential (counterclockwise angles are positive). In the inner half, fiber direction averaged 69 +/- 10 degrees, while the principal direction averaged -22 +/- 21 degrees. Therefore, while fiber and principal directions were not substantially different in the outer half, the greatest shortening occurred orthogonally to the fiber direction in the inner half. Normal and shear strains measured in a cardiac coordinate system (circumferential, longitudinal, and radial coordinates) were rotated (transformed) to "fiber" coordinates in both halves of the wall. In the outer half, normal strains observed in the fiber (-0.09 +/- 0.04) and cross-fiber (-0.04 +/- 0.04) directions were not significantly different (paired t test, p less than 0.05). In the inner half, more than twice as much strain occurred in the cross-fiber (-0.17 +/- 0.03) than in the fiber direction (-0.06 +/- 0.06). Moreover, the only shear strain that remained substantial after transformation was transverse shear in the plane of the fiber and radial coordinates. These results suggest that both reorientation and cross-sectional shape changes of myofibers or the interstitium may contribute to the large wall thickenings observed during contraction, particularly in the inner half of the ventricular wall.