Three dimensional excitation-emission fluorescence technique can be used for characterizing dissolved organic matter (DOM) in water as an innovative analysis method, which is high sensitive, simple operation, fast analysis and free sample pretreatment. The study of the composition, properties of DOM and sources of characterization methods helps people to further understand the DOM in the natural environment and the produce migration process, and to broaden its application in environmental analysis. Application basis and related environmental influencing factors of three dimensional excitation-emission fluorescence characterizing technique for dissolved organic matter were described. And the application of three dimensional excitation-emission fluorescence characterizing technique in environmental analysis was introduced, which include indirect measurement of water quality index, discrimination and tracing of organic pollutant sources and early alarming of pollution accident.