Abstract: Optical navigation sensor is a core instrument in optical autonomous navigation, the accuracy of the direction of light of the navigantion target it acquired will affect the accuracy of autonomous navigation directly. In this paper, a stepwise onorbit geometric calibration approach for optical navigation sensor is designed. Firstly, the external calibration parameters are solved. Then, the internal calibration parameters are sovled in the general camera coordinate system determined by external calibration. Because the computation source and ability is limited in the satellite, in order to using more star images to achieve estimation of calibration parameters with high accuracy, a line-by-line orthogonalization method based on least square is adopted in calibration parameter estimation. The experiments demonstrates that the on-orbit calibration approach proposed in this paper can improve the pointing accuracy of optical navigation sensor, and make it meet the requirements of optical autonomous navigation.