Isotope-labeling coupled stimulated Raman scattering (iSRS) microscopy-based high-throughput genetic screen to identify molecular mechanisms in lipid droplet incorporation of fatty acids

Saturated fatty acids (sFA) are more toxic to cells than unsaturated fatty acids (uFA). Previously, we developed a chemical imaging strategy to track the spatiotemporal dynamics of specific deuterium-labeled fatty acids in living Caenorhabditis elegans, called isotope-labeling coupled stimulated Raman scattering (iSRS) microscopy, and showed that sFAs are incorporated into lipid droplets (LDs) slower than uFAs. To study how fatty acids are integrated into LDs at different rates, we performed a high-throughput forward genetic screen using iSRS microscopy and identified mutants with enhanced sFA integration into LDs. Currently, we are sequencing these candidates to identify the molecular players in fatty acid metabolism and lipotoxicity.