We utilized Raman spectro-microscopy to non-invasively probe metabomics within single live cells, aiming to identify druggable metabolic susceptibilities from a series of patient-derived BRAF mutant melanoma cell lines. Each cell line represents a phenotype with different characteristic level of de-differentiation and BRAFi (BRAF inhibitor) resistance. First, with single-cell Raman spectroscopy and stimulated Raman scattering (SRS) microscopy, followed by transcriptomics analysis, we identified lipid processes as major metabolic functional difference between different phenotypes. We then utilized hyperspectral-SRS imaging on intracellular single organelles to identify a previously unknown susceptibility of lipid desaturation within de-differentiated cell lines. Drugging this target leads to cellular apoptosis accompanied by phase separated intracellular domains. The integration of subcellular Raman spectro-microscopy with lipidomics and transcriptomics suggests highly heterogenous metabolic responses and possible lipid regulatory mechanisms underlying this pharmacological treatment. Our method should provide a general approach in spatially-resolved single cell metabolomics studies.
Innovations in optical spectroscopy and microscopy have revolutionized our understanding in
biological systems at sub-cellular levels. In this talk, I will discuss about our recent development by coupling stimulated Raman scattering (SRS) microscopy with chemical probes that could allow new subcellular bioanalysis in live cells. The introduced tags offer additional SRS contrast channel for quantification of biological contents that were previously difficult. Both physical and chemical principles underlying the optical microscopy will be presented, as well as our efforts in biomedical applications including cancer- and neuronal- metabolism.
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