Ambient high-spatial-resolution mass spectrometry imaging: recent advances in technologies and metabolomics studies

Achieving high-spatial-resolution in ambient mass spectrometry imaging (MSI) is critical in accurately characterizing the spatial distribution of biomolecules. Ambient MSI enables in situ mapping spatial heterogeneity of metabolites under ambient conditions, facilitating molecular imaging from multicellular structure down to subcellular level. In this review, we first summarized the strategies developed to enhance spatial resolution in ambient MSI, including technological innovations, advances in sample preparation, and data-driven approaches. We then discussed the applications of these techniques in metabolomics studies and addressed current challenges and limitations. Finally, we highlighted emerging opportunities and future directions for achieving ambient high-spatial-resolution MSI.