Laser induced fluorescence (LIF) is a non-intrusive optical technique that uses fluorescent dyes to measure whole-field fluid scalars such as temperature, concentration, pH, etc. LIF measurements’ accuracy is strongly influenced by the fluorescent dye's behavior under different experimental conditions. In particular, ratiometric LIF thermometry accuracy depends on the correct selection of fluorescent dyes mixtures. Therefore, a thorough characterizations of fluorescent dyes is needed to obtain optimal mixtures and suitable optical configurations for given experimental conditions. This work presents the experimental characterization of fluorescein-27 (FL27) and rhodamine-B (RhB) mixtures to determine suitable aqueous solutions for ratiometric LIF thermometry. The mixtures' fluorescence emission intensity was measured with a spectrofluorometer, and the influence of concentration ratio (\documentclass[12pt]{minimal}\begin{document}$C_{\text{RhB}}/C_{\text{FL27}}$\end{document}CRhB/CFL27), temperature, excitation wavelength (λext), and pH were analyzed. The results show that the temperature dependence of FL27 emission intensity changed from a negative to a positive value as the excitation wavelength increased. The temperature sensitivity (4.0% per °C) of RhB and FL27 mixture under 532 nm excitation wavelength was found to be higher than that of the commonly used mixture of RhB and Rh110 (2.0% per °C) at the same excitation wavelength. While the emission intensities of the dyes are sensitive to pH value, the temperature dependence is unaffected. The influence of concentration ratio on temperature sensitivity depends on both the detected bands of the emitted spectrum and the temperature; the concentration ratio should be selected based on the measured temperature scope. A new multicolor method or advanced two color method with high temperature sensitivity (6.0% or 10.0% per °C) is presented. This technique was specially developed to improve whole-field temperature measurements.