Laser tweezers Raman spectroscopy (LTRS) integrating with optical tweezers with confocal Raman spectroscopy ensures the trapping, manipulation, measurement and biochemical analysis under physiological conditions for the single living cell analysis in suspension. Due to the unique advantage in single-cell research, LTRS has drawn more and more attention. In this review, the biomedical applications and the development of laser tweezers Raman spectroscopy in recent years are taken as the main line, centering around the four areas: Raman spectroscopy in the analysis of cell and cell sorting, multi-optical tweezers Raman spectroscopy systems, micro-fluidic transport systems and the technology of multi-mode analysis, to summarize the recent development of LTRS in multi-data acquisition and integration, high efficiency, high-throughput and the transport of automation by combination with other novel techniques or upgrading technologies. At the same time, the technology applied in biomedicine is introduced. Finally, a perspective of the future development for the powerful cell analysis technology is presented.