In vivo surface-enhanced Raman scattering techniques: nanoprobes, instrumentation, and applications

Surface-enhanced Raman scattering (SERS) has emerged as a powerful tool in various biomedical applications, including in vivo imaging, diagnostics, and therapy, largely due to the development of near-infrared (NIR) active SERS substrates. This review provides a comprehensive overview of SERS-based applications in vivo, focusing on key aspects such as the design considerations for SERS nanoprobes and advancements in instrumentation. Topics covered include the development of NIR SERS substrates, Raman label compounds (RLCs), protective coatings, and the conjugation of bioligands for targeted imaging and therapy. The review also discusses microscope-based configurations such as scanning, widefield imaging, and fiber-optic setups. Recent advances in using SERS nanoprobes for in vivo sensing, diagnostics, biomolecule screening, multiplex imaging, intraoperative guidance, and multifunctional cancer therapy are highlighted. The review concludes by addressing challenges in the clinical translation of SERS nanoprobes and outlines future directions, emphasizing opportunities for advancing biomedical research and clinical applications.