Deuterium-oxide-assisted stimulated Raman scattering microscopy: metabolic imaging for precision medicine

Metabolomics is transforming personalized medicine by enabling the customization of treatment plans based on the unique metabolic profiles of individual patients. However, general metabolomics technologies are destructive and lack spatial resolution, which limits their application in live-cell analysis and dynamic imaging studies. Furthermore, these methods typically lack sufficient spatial resolution, which limits their capacity to capture metabolic heterogeneity at the single-cell or subcellular level. These limitations hinder the study of spatially and temporally dynamic metabolic processes in diseases such as cancer, metabolic, and neurological disorders.

Our review summarizes advances in deuterium-oxide-assisted stimulated Raman scattering ( $D_{2}O\text{-}\mathrm{SRS}$ ) microscopy, with a focus on its capabilities for real-time, high-resolution metabolic imaging. We discuss how this technology enables real-time tracking of biosynthetic processes in living systems, contributing to the advancement of precision diagnostics and therapeutic strategies, highlighting its recent progress in tumor metabolism monitoring, and drug sensitivity testing for infectious diseases, as well as its future potential applications.

We outline the nonlinear optical principles of SRS, the deuterium oxide ( $D_{2}O$ ) labeling strategy that generates carbon-deuterium (C-D) vibrational signals in newly synthesized macromolecules, and key applications in tracking lipid, protein, and nucleic acid biosynthesis.

$D_{2}O\text{-}\mathrm{SRS}$ enables real-time, quantitative imaging of metabolic activity with subcellular resolution and minimal perturbation, offering chemical and spatial insight into metabolic dynamics.

$D_{2}O\text{-}\mathrm{SRS}$ microscopy represents a significant advancement in live metabolic imaging, offering a powerful tool for investigating disease-related metabolic dysfunction. This technology enables high-resolution, real-time metabolic imaging in living systems. As a result, $D_{2}O\text{-}\mathrm{SRS}$ is expected to promote the transformation and implementation of basic metabolic research into clinical applications.