InSitu Untargeted Metabolite Identification and Spatial Mapping by Clustering Resampling Mass Spectrometry Imaging

However, <i>in situ</i> untargeted metabolite identification remains challenging due to the inherent trade-off between spatial resolution and <i>in situ</i> tandem mass spectrometry (MS2) acquisition. Here, we developed clustering resampling mass spectrometry imaging (CR-MSI), a method that innovatively converts <i>in situ</i> MS2 acquisition from countless pixel scans to a few representative points testing.

CR-MSI allows the acquisition of both spatial localization and structural fragmentation data for metabolites from a single tissue section. Additionally, we have engineered an integrated device that automates the CR-MSI workflow, transforming MS2 acquisition from an empirical, operator-dependent procedure to an objective, data-driven process. Moreover, the synchronization between MSI data acquisition and processing has improved the efficiency and overall MSI throughput.