Abstract: Three-dimensional fluorescence spectrum is developed as a kind of new fluorescence analysis technique since 1980s. Comparing to the ultraviolet fluorescence features,it could be obtained the fluorescence intensities of excitation wavelength and emission wavelength when changing at the same time for three-dimensional fluorescence spectrum,which is used for qualitative and quantitative analysis of multicomponent mixtrue and environment, biology and chemistry. However,it is still merely applicated in the testing and research of gemstones. 25pieces of the diamond samples from Mengyin,Shandong Province,are tested and studied by the three-dimensional fluorescence and ultraviolet fluorescence. The results show that the three-dimensional fluorescence spectrum features of the diamond samples could be divided into four types,namingly non-fluorescence spectrum peak,one fluorescence spectrum peak,double fluorescence spectrum peaks and three fluorescence spectrum peaks,which are well with the ultraviolet fluorescence features.