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Here, we present tenfold expansion mass spectrometry imaging (10X ExMSI), an expansion MALDI\u2010MSI workflow that reaches an effective lateral resolution of \u223c500\u00a0nm on standard commercial instruments using a 5\u00a0\u00b5m acquisition step size without pixel oversampling. The SDS\u2010free digestion protocol minimizes lipid leaching and supports broad detection of major lipid classes, including glycerophospholipids and sphingolipids, although a reduction in primary amine\u2010containing lipids is observed due to cross\u2010linking. Using 10X ExMSI, we resolve subcellular structures in mouse brain tissues, such as dendritic arborizations, that are challenging to access with existing MALDI\u2010MSI implementations. We further demonstrate expansion\u2010based MSI on cultured A549 cells, achieving subcellular\u2010level lipid mapping. The method is fully compatible with standard MALDI\u2010MSI systems, providing an accessible and scalable route to high\u2010resolution, label\u2010free molecular imaging. These capabilities open new opportunities for studying cellular architecture and molecular heterogeneity in biological and biomedical research.<\/jats:p>","DOI":"10.1002\/anie.1256471","type":"journal-article","created":{"date-parts":[[2026,8,7]],"date-time":"2026-08-07T12:59:36Z","timestamp":1786107576000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Tenfold Expansion Submicrometer\u00a0MALDI Mass Spectrometry Imaging of Tissues and Cultured Cells"],"prefix":"10.1002","volume":"65","author":[{"given":"Chengyi","family":"Xie","sequence":"first","affiliation":[{"name":"Department of Chemistry Hong Kong Baptist University  Hong Kong China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9294-2809","authenticated-orcid":false,"given":"Jianing","family":"Wang","sequence":"additional","affiliation":[{"name":"School of Marine Sciences Hainan University  Haikou 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