Crime scene investigation relies heavily on evidence derived from biological materials like bodily fluids, tissues, hair, and fingermarks. It is possible to obtain a great deal of important information about the crime and the donor individuals from the biological materials. In recent years, vibrational spectroscopy has emerged as a rapid, non-destructive, label-free, and user-friendly technique for biological analysis. When it comes to forensic biological analysis, it is most frequently applied to bodily fluids. This chapter discusses recently published studies in which vibrational spectroscopy coupled with chemometric algorithms was employed for forensic bodily fluid investigation, such as identification of bodily fluids, determination of the donor’s species, estimation of the deposition time, and characterization of the donor’s basic information (e.g., age, sex, race). Problems encountered in applying these techniques to routine forensic practice and perspectives with regard to the further improvement of forensic bodily fluid spectral investigation techniques are also discussed in this chapter.