Bridging the molecular resolution gap: integrating imaging mass cytometry with mass spectrometry for deeper insights into inflammation 4621

Abstract Description Emerging single-cell spatial-omics techniques, such as Imaging Mass Cytometry (IMC), provide high-resolution cellular phenotyping in heterogeneous tissues. However, these methods are limited in detecting molecular features like lipids, glycans, and metabolites. In contrast, mass spectrometry imaging (MSI) excels at analyzing these features but lacks the cellular resolution needed for precise single cell spatial context. To bridge this gap, we developed an integrative framework combining IMC and MSI through a specialized wet lab workflow and refined image alignment software. This approach enabled us to investigate glycosylation changes in human liver inflammation. While individual technologies detected only minor, previously documented changes, integrating their data revealed distinct single-cell molecular signatures, including unique N-glycan profiles in inflammatory immune cells. These profiles correlated with macrophage markers such as CD206, CD163, and HLA-DR. Spatial transcriptomics further validated these findings. Ongoing studies extend this work to animal models, leveraging mass spectrometry to further characterize immune cell glycosylation at the single-cell level. By integrating spatial-omics technologies, we uncover intricate molecular glycosylation patterns of individual cells in complex tissues, opening new frontiers for understanding biology and advancing therapy. Funding Sources RSG-24-1255025-01-IBCD and R01CA258882 to S.G.. Topic Categories Technological Innovations in Immunology (TECH)