The spectroscopic identification of body fluids in situ is a major objective in forensic science. This approach offers the confirmatory, nondestructive, rapid, and on-scene identification of various body fluids. Although Raman spectroscopy has shown tremendous promise toward this goal in prior proof-of-concept experiments, a significant challenge which still remains is substrate interference. The current project prepared simulated semen evidence on skin, glass, and various fabrics. Raman data were accumulated from stains without any pretreatment using a common confocal mapping spectrometer using 785 nm laser excitation. The results demonstrate that the spectroscopic interferences encountered by substrates can be reduced and eliminated by using a combination of existing subtraction techniques and chemometric models. Heterogeneous substrates proved most challenging; however, automatic subtraction treatment and location of fluid hotspots produced a clear spectroscopic signature of semen in every instance. (Publisher abstract modified)
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