Biplane fluoroscopy is used for dynamic in vivo three-dimensional motion analysis of various joints of the body. Cross-scatter between the two fluoroscopy systems may limit tracking accuracy. This study measured the magnitude and effects of cross-scatter in biplane fluoroscopic images. Four cylindrical phantoms of 4-, 6-, 8-, and 10-in. diameter were imaged at varying kVp levels to determine the cross-scatter fraction and contrast-to-noise ratio (CNR). Monte Carlo simulations quantified the effect of the gantry angle on the cross-scatter fraction. A cadaver foot with implanted beads was also imaged. The effect of cross-scatter on marker-based tracking accuracy was investigated. Results demonstrated that the cross-scatter fraction varied from 0.15 for the 4-in. cylinder to 0.89 for the 10-in. cylinder when averaged across kVp. The average change in CNR due to cross-scatter ranged from 5% to 36% CNR decreases for the 4- and 10-in. cylinders, respectively. In simulations, the cross-scatter fraction increased with the gantry angle for the 8- and 10-in. cylinders. Cross-scatter significantly increased static-tracking error by 15%, 25%, and 38% for the 6-, 8-, and 10-in. phantoms, respectively, with no significant effect for the foot specimen. The results demonstrated submillimeter marker-based tracking for a range of phantom sizes, despite cross-scatter degradation.