Abstract
Molecular biomarkers play an essential role in accurate disease diagnosis and personalized treatment. Dysregulated metabolism is closely associated with disease development and progression. The discovery of metabolic biomarkers could remarkably promote precision diagnosis and personalized treatment. Current metabolomics approaches can profile a large number of metabolites but are primarily destructive and lack sufficient spatial resolution, which hinders quantitative measurements of the highly dynamic and heterogeneous intracellular metabolic processes. This further limits the discovery of metabolic biomarkers in these diseases. Stimulated Raman scattering (SRS) microscopy addresses these gaps by enabling label-free imaging with high sensitivity, molecular specificity, and subcellular resolution. Integrating Raman-active vibrational probes further extends this approach, allowing for real-time tracking of low-abundance biomolecules and metabolic processes. These capabilities have enabled the discovery of biomarkers for disease diagnosis. In this review, we focus on recent advancements in SRS imaging technologies and data analysis methods and their applications in biomarker discovery and precision medicine. Furthermore, future perspectives and emerging trends in this rapidly evolving research area are discussed.