Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030, United states ?Program in Developmental Biology, Baylor College of Medicine, Houston, Texas 77030, United states of america Gladstone Institute of Cardiovascular Disease, Division of Biochemistry and Biophysics, University of California, San Francisco, California 94158, United states of america Diabetes and Endocrinology Study Center and Department of Medicine, Baylor College of Medicine, Houston, Texas 77030, United StatesS * Supporting InformationABSTRACT: Metabolic fingerprinting delivers precious information and facts around the physiopathological states of cells and tissues. Regular imaging mass spectrometry and magnetic resonance imaging are unable to probe the spatial-temporal dynamics of metabolites in the subcellular level because of either lack of spatial resolution or inability to carry out live cell imaging. Here we report a complementary metabolic imaging approach that’s primarily based on hyperspectral stimulated Raman scattering (hsSRS). We demonstrated the usage of hsSRS imaging in quantifying two important neutral lipids: cholesteryl ester and triacylglycerol in cells and tissues. Our imaging final results revealed previously unknown changes of lipid composition related with obesity and steatohepatitis. We further made use of stable-isotope labeling to trace the metabolic dynamics of fatty acids in reside cells and reside Caenorhabditis elegans with hsSRS imaging. We located that unsaturated fatty acid has preferential uptake into lipid storage whilst saturated fatty acid exhibits toxicity in hepatic cells. Simultaneous metabolic fingerprinting of deuterium-labeled saturated and unsaturated fatty acids in living C. elegans revealed that there’s a lack of interaction involving the two, in contrast to previously hypothesized. Our findings offer new approaches for metabolic tracing of neutral lipids and their precursors in living cells and organisms, and could potentially serve as a common method for metabolic fingerprinting of other metabolites.INTRODUCTION Understanding the complex metabolic processes that occur within living organisms offers critical pathways to tackle key healthcare challenges such as diabetes and cancer. In the human body, each sort of cell or tissue includes a special “metabolic fingerprint” that characterizes its specific function. The emerging field of metabolomics aims to uncover metabolic fingerprints of tissues at various physiopathological states.1286754-61-7 Chemscene Its progress relies heavily on technological renovations with evolving capability of detecting and quantifying the thousands of metabolites (also referred to as metabolome) to be found within a biological sample.150730-41-9 Price 1 A wide range of mass spectrometry (MS) approaches (generally coupled to gas chromatography or liquid chromatography) have been employed to characterize the metabolome owing to their higher sensitivity and specificity.PMID:33529425 To date, MS remains the essential platform that is certainly applied to examine relative metabolite profile differences among biological?2014 American Chemical Societysamples. Nevertheless, it only offers a snapshot of metabolite profile at a particular time point, and typically devoid of any spatial context. Current developments in imaging mass spectrometry (IMS) offer the much-needed spatial data for understanding disease mechanisms and their progresses. When combined with isotope labeling, IMS is extremely strong in characterizing metabolic fate of tiny molecules at higher resolution.2,three Magnetic resonance imaging (MRI) is a further strategy th.