In vivo analyses of pacemaker lead motion during the cardiac cycle have become important due to incidences of failure of some of the components. For the calculation and evaluation of in vivo stresses in pacemaker leads, the 3D motion of the lead must be determined. To accomplish this, we have developed a technique for calculation of the overall and relative 3D position, and thereby the 3D motion, of in vivo pacemaker leads through the cardiac cycle.Biplane image sequences of patients with pacemakers were acquired for at least two cardiac cycles. After the patient acquisitions, biplane images of a calibration phantom were obtained. The biplane imaging geometries were calculated from the images of the calibration phantom. Points on the electrodes and the lead centerlines were indicated manually in all acquired images. The indicated points along the leads were then fit using a cubic spline. In each projection, the cumulative arclength along the centerlines in two temporally adjacent images was used to identify corresponding points along the centerlines. To overcome the non-synchronicity of the biplane image acquisition, temporal interpolation was performed using these corresponding points based on a linear scheme. For each time point, corresponding points along the lead centerlines in the pairs of biplane images were identified using epipolar lines. The 3D lead centerlines were calculated from the calculated imaging geometries and the corresponding image points along the lead centerlines. From these data, 3D lead motion and the variations of the lead position with time were calculated and evaluated throughout the cardiac cycle. The reproducibility of the indicated lead centerlines was approximately 0.3 mm. The precision of the calculated rotation matrix and translation vector defining image geometry were approximately 2 mm. 3D positions were reproducible to within 2 mm. Relative positional errors were less than 0.3 mm. Lead motion correlated strongly with phases of the cardiac cycle. Our results indicate that complex motions of in vivo pacemaker leads can be precisely determined. Thus, we believe that this technique will provide precise 3D motion and shapes on which to base subsequent stress analysis of pacemaker lead components.