In this paper, a new on-orbit polarization calibration method for the multichannel polarimetric camera is presented. A polarization calibration model for the polarimetric camera is proposed by taking analysis of the polarization radiation transmission process. In order to get the polarization parameters in the calibration model, an on-orbit measurement scheme is reported, which uses a solar diffuser and a built-in rotatable linear analyzer. The advantages of this scheme are sharing the same calibration assembly with the radiometric calibration and acquiring sufficient polarization accuracy. The influence of the diffuser for the measurement is analyzed. By using a verification experiment, the proposed method can achieve on-orbit polarization calibration. The experimental results show that the relative deviation for the measured degree of linear polarization is 0.8% at 670 nm, which provides a foundation for the accurate application of polarimetric imaging detection.