The current scientific world majorly focuses on nanobiotechnology, biomedical studies, and molecular markers for endocrine, cardiovascular, and various oncological diseases. Various biomolecules, such as carotenoids and lipids, are being studied in the area of biotechnology. Raman spectroscopy (RS) has developed to be a crucial method for analyzing different biological techniques at the tissue and cellular level in the field of biotechnology and medicine. RS is used for the analysis of medical, biological, photosynthetic, analytical, and algal research. Different bacteria and their chemical composition can also be detected using RS. Different types of RS are used to analyze the composition of the cell. Tissue selection is a crucial process while analyzing a sample using RS. This chapter summarizes different methods used to analyze the sample using RS. The advantage and disadvantages of different methods using RS are also described in this chapter.