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In order to help the undergraduate students to acquire a better understanding of the principles of the fluorescence analysis and improve their comprehensive analysis and problem-solving skills, the content and teaching process of the Vitamin B2 fluorescence experiment were redesigned. The basic theory was intensified through the identifications of the Raman and Rayleigh scatterings ； the functions of the component parts of the instrument were better comprehended by adjusting parameters like the pass band, high voltages of photomultiplier tube and so on. Through the well-designed question and answer session with specific data and experimental operation, the learning initiative and creativity of the students were fully developed. The ability of analyzing and solving problems was increased and the teaching quality of the fluorescence experiment was effectively improved indeed. [ABSTRACT FROM AUTHOR]