Purpose:Determine systematic deviations between 2D/3D and 3D/3D image registrations with six degrees of freedom (6DOF) for various imaging modalities and registration algorithms on the Varian Edge Linac.Methods:The 6DOF systematic errors were assessed by comparing automated 2D/3D (kV/MV vs. CT) with 3D/3D (CBCT vs. CT) image registrations from different imaging pairs, CT slice thicknesses, couch angles, similarity measures, etc., using a Rando head and a pelvic phantom. The 2D/3D image registration accuracy was evaluated at different treatment sites (intra‐cranial and extra‐cranial) by statistically analyzing 2D/3D pre‐treatment verification against 3D/3D localization of 192 Stereotactic Radiosurgery/Stereotactic Body Radiation Therapy treatment fractions for 88 patients.Results:The systematic errors of 2D/3D image registration using kV‐kV, MV‐kV and MV‐MV image pairs using 0.8 mm slice thickness CT images were within 0.3 mm and 0.3° for translations and rotations with a 95% confidence interval (CI). No significant difference between 2D/3D and 3D/3D image registrations (P>0.05) was observed for target localization at various CT slice thicknesses ranging from 0.8 to 3 mm. Couch angles (30, 45, 60 degree) did not impact the accuracy of 2D/3D image registration. Using pattern intensity with content image filtering was recommended for 2D/3D image registration to achieve the best accuracy. For the patient study, translational error was within 2 mm and rotational error was within 0.6 degrees in terms of 95% CI for 2D/3D image registration. For intra‐cranial sites, means and std. deviations of translational errors were −0.2±0.7, 0.04±0.5, 0.1±0.4 mm for LNG, LAT, VRT directions, respectively. For extra‐cranial sites, means and std. deviations of translational errors were ‐ 0.04±1, 0.2±1, 0.1±1 mm for LNG, LAT, VRT directions, respectively. 2D/3D image registration uncertainties for intra‐cranial and extra‐cranial sites were comparable.Conclusion:The Varian Edge radiosurgery 6DOF‐based system, can perform 2D/3D image registration with high accuracy for target localization in image‐guided stereotactic radiosurgery.The work was supported by a Research Scholar Grant, RSG‐15‐137‐01‐CCE from the American Cancer Society.