Raman spectroscopy which belongs to scattering spectroscopy obtained molecular vibrational and rotational information to achieve detection and analysis of molecular structure and corresponding changes through recording the frequency shift when light interacted with materials. Compared with routine biochemical analysis, Raman spectroscopy has the advantage of non-invasive, label-free and no sample requirement. Raman spectroscopy has been widely applied in biomedical field such as human tissue, organs, cells and human body fluids for disease diagnosis. This article mainly focuses on recent research advances of Raman spectroscopy in human semen. Firstly, Raman spectroscopy(including surface-enhanced Raman spectroscopy, SERS) employed in forensic science for semen analysis, and some related data processing methods were introduced, then Raman spectroscopy involved investigations of male fertility was highlighted, more specifically, the Raman-based qualitative and quantitative analysis which assist the objective detection and evaluation of male fertility. Furthermore, studies of single sperm cell based on micro-Raman system to characterize and evaluate sperm quality and the preliminarily obtained Raman biomarkers which indicate high-quality sperm cell were introduced. Finally, the potential development of Raman spectroscopy involved in reproduction and fertility field was also discussed.