Binocular stereoscopic vision can be used for space-based space targets near observation. In order to solve the problem that the traditional binocular vision system cannot work normally after interference, an online calibration method of binocular stereo measuring camera with self-reference is proposed. The method uses an auxiliary optical imaging device to insert the image of the standard reference object into the edge of the main optical path and image with the target on the same focal plane, which is equivalent to a standard reference in the binocular imaging optical system; When the position of the system and the imaging device parameters are disturbed, the image of the standard reference will change accordingly in the imaging plane, and the position of the standard reference object does not change. The camera's external parameters can be re-calibrated by the visual relationship of the standard reference object. The experimental results show that the maximum mean square error of the same object can be reduced from the original 72.88mm to 1.65mm when the right camera is deflected by 0.4 degrees and the left camera is high and low with 0.2° rotation. This method can realize the online calibration of binocular stereoscopic vision measurement system, which can effectively improve the anti - jamming ability of the system.