Direct imaging of metabolism at sub-cellular resolution provides insights to understand single-cell physiology at spatially resolved manner. Here, we presented a metabolic profiling platform based on Optical Photothermal Infrared (OPTIR) spectroscopic imaging that enables in situ lipid metabolism characterization at sub-cellular resolution. We applied the developed metabolic imaging platform to study lipid synthesis from azide-labeled fatty acids across various systems including neuroglioma cells, human-induced pluripotent stem cells (iPSCs), iPSC-derived microglia cells, and iPSC-derived brain organoids. The specificity of the developed platform was further improved by coupling a fluorescence module that enables cell type specific-metabolic profiling. Collectivity, the proposed metabolic imaging platform opens a vast potential for in situ metabolism characterization at the single-cell level.
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