Unlocking Lipid Spatialomics in FFPE Tissues via MALDI Mass Spectrometry Imaging : Lipid MS/MS raw files

Unlocking Lipid Spatialomics in FFPE Tissues via MALDI Mass Spectrometry Imaging Lipid annotations were confirmed by tandem mass spectrometry (MALDI-MS/MS) on the timsTOF fleX operated in qTOF mode, in both positive- and negative-ion polarity. DHB-coated sections previously used for imaging were reused for targeted MS/MS, enabling direct correlation between ion localization and fragmentation data. Precursor ions of interest were selected from imaging datasets based on spatial distribution and intensity, isolated with a 0.1 Da window, and fragmented by collision-induced dissociation (CID) with nitrogen at 30–45 eV, using a laser repetition rate of 10,000 Hz and 500 laser shots per spectrum, over an m/z range of 50–1350.