Multiplexed mass spectrometric lipid metabolic tracing with single cell sensitivity
Lipid metabolic tracing is a key technology to understand lipid metabolism in health and disease. We have previously demonstrated tracing with picomole sensitivity and lipid class resolution using alkyne-labeled lipids and detection by fluorogenic click reaction combined with TLC separation. To take advantage of mass spectrometry for analysis, we have developed a family of novel click chemistry MS reporter molecules that combine highly improved sensitivity with enhanced specificity. Introduction of heavy isotopes into the reagents allows multiplexed analysis that saves time and strongly reduces technical noise. The presentation will demonstrate the performance of the system in tracing hepatic lipid metabolism in more than ten lipid classes with several hundreds labeled lipid species. Finally, I will demonstrate how the femtomole sensitivity of the method can be used to trace fatty acid incorporation in single hepatocytes.
Rheinische Friedrich-Wilhelms-Universität, Bonn, Allemagne