Raman spectroscopy was shown to be a panacea for identifying body fluids, marking a watershed event in the field of serology. Raman spectra of certain fluids can be used for more than just identification; they can also be analyzed using chemometrics to learn things like the fluid's origin, the donor's race and gender, the donor's age, and the amount of time since depositing. This study aimed to determine the ability of the Raman Spectroscopy to identify the semen-blood mixture. Subjects & Methods: The present work is using Raman Spectroscopy to identify semen-blood mixture samples. Fifteen body fluids samples were obtained, and mixed together. All of these samples were examined using a 3D confocal Raman imaging microscope with a 50x long-range objective (numerical aperture = 0.75) and a WiTec alpha300 R confocal Raman spectrometer. Results: We found that the average Raman spectra of dry traces of semen-blood mixture constitutes mainly of hemoglobin, choline, and spermine phosphate hexahydrate. This proof of concept approach shows the potential for Raman spectroscopy to identify semen mixtures with blood during forensic analysis. Conclusion: Different mixtures of semen with blood can be distinguished from each other using Raman spectroscopy couple with chemometrics.