In order to test and analyze the fluorescence life of up-conversion nanoparticles (UCNPs), a system was designed for high-speed acquisition and measurement of fluorescence signal data of UCNPs. The system programmed the virtual original fluorescent signals through MATLAB, completed the filtering and data fitting, embeded the data program script into LabVIEW for calculating and processing, and displayed the fluorescent signal waveform, fitting data waveform, fluorescence number, fluorescence life and extreme of the life on the host computer. The research results show that the system software simulation and numerical calculation simulation have achieved basically consistent results, the error between the two is less than 3%, with good stability and accuracy. This research work has great guiding value for the development of UCNPs fluorescence lifetime measurement system.