This paper explores the feasibility of non-destructive extraction of urine footprints on rough paper by visible laser and quantitatively comparing the effects of footprint development. Three kinds of visible lasers and 11 kinds of visible band filters were used to take visible laser fluorescence photography of urine footprints on 4 categories of 20 kinds of common types of paper to observe the effect of footprints. The Matlab program was created, and gray-scale fluorescence luminance standard deviation index was introduced to compare the degree of footprint visualization. The study found that the band combinations of the 20 samples with the best development effect were almost different, but the 445 nm blue laser was used as the excitation light, and the 570 nm receiving filter was used to develop the urine footprints of all samples to the maximum extent, the detection rate of urine footprints was 100%. In addition to the urine footprint samples on black printer paper and newspaper with complex color background, the effective visualization rate of urine footprint samples was 90%. The results show that visible laser fluorescence photography can be used to visualize urine footprints on rough paper, and the standard deviation of gray-scale fluorescence luminance can be used to measure the degree of urine footprints development.