The traditional method of establishing the relationship between fluorescence brightness and dye concentration through concentration gradient, and evaluating the sensitivity of imaging detection is too repetitive and cumbersome, and is obviously affected by handwork. Therefore, the reliability and reproducibility of the data are low. This work provides a standard concentration and imaging luminance relationship curve for equipment manufacturers through concentration preparation and detection quantification by authoritative institutions based on indocyanine green dye. It only needs to detect the excitation light power and simulated fluorescence radiation of the equipment in the imaging system, and find the relationship curve to obtain the relationship between "radiation and concentration" and "signal-to-background ratio-concentration", which can be used as a quantitative basis for the camera system. All operations can achieve traceability operation, which greatly reduces the human experimental error, and provides reliable evidence for the detection sensitivity evaluation of bioimaging system based on fluorescent dyes.