This study proposed a method to calibrate tube focus spot and the center plane of rotation in computed tomography system. In the method, the tube was rotated to 0° and 180° respectively, and then one metal jig with symmetric windows A and B was scanned at each position under the tube cool and static condition. According to the geometry of tube focus spot, aperture center of the collimator and jig, the distance between tube focus spot and the center plane of rotation were calculated with the X ray transmittance data after denoising, mean value and normalization. To verify the practicability and validity of the method, the tube focus spot in a 16 slices CT system (Brivo CT385, GE, China) was calibrated, and the result after calibration was validated by scanning a polaroid film. The validation result showed that the deviation between tube focal spot and center plane of rotation was 0.02 mm and was in the error range within ± 0.1 mm. The results of this study showed that, as a simple and low-cost design, the method could be used for fast calibration between tube focus spot and the center plane of rotation.