In order to analyze, identify and trace the sources of pollutants in water bodies, the use of three-dimensional fluorescence spectroscopy to determine the types and contents of pollutants in water has the advantages of high accuracy and fast determination. The application of three-dimensional fluorescence spectroscopy in the analysis of organic pollutants in water bodies has been studied. Through the characterization of the characteristic fluorescence spectra of different organic pollutants in sewage, the identification and traceability of the pollutants can be quickly and efficiently realized. The current commonly used three-dimensional fluorescence fingerprint analysis methods are introduced, and the correlation between the fluorescence properties of DOM in sewage and organic pollution indicators are discussed. The application status of three-dimensional fluorescence spectrum analysis technology in sewage treatment is discussed, and its application prospects in the field of sewage treatment are predicted.