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Background Accurate camera and hand‐eye calibration are essential to ensure high‐quality results in image‐guided surgery applications. The process must also be able to be undertaken by a nonexpert user in a surgical setting. Purpose This work seeks to identify a suitable method for tracked stereo laparoscope calibration within theater. Methods A custom calibration rig, to enable rapid calibration in a surgical setting, was designed. The rig was compared against freehand calibration. Stereo reprojection, stereo reconstruction, tracked stereo reprojection, and tracked stereo reconstruction error metrics were used to evaluate calibration quality. Results Use of the calibration rig reduced mean errors: reprojection (1.47 mm [SD 0.13] vs. 3.14 mm [SD 2.11], p‐value 1e−8), reconstruction (1.37 px [SD 0.10] vs. 10.10 px [SD 4.54], p‐value 6e−7), and tracked reconstruction (1.38 mm [SD 0.10] vs. 12.64 mm [SD 4.34], p‐value 1e−6) compared with freehand calibration. The use of a ChArUco pattern yielded slightly lower reprojection errors, while a dot grid produced lower reconstruction errors and was more robust under strong global illumination. Conclusion The use of the calibration rig results in a statistically significant decrease in calibration error metrics, versus freehand calibration, and represents the preferred approach for use in the operating theater.

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