In view of the uncertainty in the calibration process of two-color plane laser-induced fluorescence (PLIF) temperature measurement, a new calibration method is proposed, in which the influence of fluorescence yield is considered. The calibration process was carried out at high and low temperature region, respectively. Then, the bias of thermometry results origin from quenching is restrained. This new calibration method is validated in a jet flame with temperature range of 1300–1800 K. Here, the temperature results from Coherent Anti-Stokes Raman scattering (CARS), single-point calibrated PLIF, and two-point calibrated PLIF are all acquired with the maximum standard errors of 13 K, 36 K, and 37 K, respectively. The temperature deviation between the average results from PLIF and Coherent Anti-Stokes Raman scattering (CARS) is 120 K and 10 K, when the two-point and one-point calibration methods are used. Therefore, the two-point calibrated PLIF is preferred in the combustion field, especially with a large temperature range and strong quenching coefficient.