 This paper proposes a new method for evaluating the performance of bioluminescent probes. By simultaneously measuring the photoacoustic signal for the non-radiative energy process and the fluorescence for the radiative energy process, it is possible to gain more accurate and detailed information about the probe's behavior in complex environments. This method has been successfully applied to evaluate the performance of a bioluminescent probe in various environments, demonstrating its effectiveness in improving the accuracy of quantifying the probe's activity. This article was authored by Huang Xiong Chen, Xiaoyutang, Huang Shui Nian, and others.