Recently, OH planar laser-induced fluorescence (PLIF) using the broadband, ultrashort femtosecond-duration (fs-duration) and the thermally assisted vibrational transfer in excited state has been investigated in flames. In this present work, we first measured temperature by thermally assisted OH laser-induced fluorescence (TALF) method with a single ultrashort broadband fs laser. In the experiment, the fs excitation of OH at ultraviolet wavelength is followed by fluorescence detection from two different vibrational bands. The ratio of two measured (1–0) and (0–0) band fluorescence is calibrated with calculated temperature using Chemkin PRO PRIMIX. The calibrated results are used in measuring temperature distributions in different laminar flames. It is found that TALF method using the fs laser can detect 2D temperature distribution in the burnt area with high OH fluorescence signal. However, OH chemiluminescence brings inevitable noise at the flame front that the TALF method does not perform well. And because (1–0) band fluorescence is so weak, the noise from the camera sensor and imaging intensifier (I.I.) remains at the measured temperature imaging. In conclusion, quantitative temperature measurement based on OH TALF based on a single broadband, ultrashort fs laser can be applied in laminar flames with high frequency by a simple experiment setup.