This paper explores the characteristics of absorption and fluorescence spectra of p-xylene within the temperature range that is frequently encountered during the mixture formation in internal combustion engines. At 266 nm wavelength, the p-xylene absorption cross section shows a mean value of (3.4 ± 0.2) × 10−19 cm2/molecule within the temperature range from 423 to 623 K in N2. As expected, fluorescence peak intensity decreases by a factor of 3 when the temperature increases by 100 K, due to a increasing non-radiative decay rate of excited state at increasing temperatures. In addition, the suitability of p-xylene for temperature measurements in the gas phase via the single-wavelength excitation (at 266 nm) two-color detection laser-induced fluorescence imaging is explored. Combinations of spectral detection bands were compared and the combination of 320/289 nm provides the best temperature performance with a relative error of 2.6% within the investigated temperature range. It is also shown that the temperature field measurement has not been strongly affected by the laser attenuation.