{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T02:43:37Z","timestamp":1792032217191,"version":"4.4.0"},"reference-count":51,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,10,1]],"date-time":"2025-10-01T00:00:00Z","timestamp":1759276800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005892","name":"China Academy of Chinese Medical Sciences","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100005892","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>Leukemia, a heterogeneous group of hematological malignancies, is characterized by abnormal proliferation of immature hematopoietic cells. Current diagnostics primarily rely on morphological evaluation for subtype classification, methods that are subjective and labor-intensive. To overcome these limitations, a High-Content Spectral Raman Pathology Imaging platform (H-SRPI) was introduced.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>H-SRPI imaging enables profiling of proteins, nucleic acids, saturated and unsaturated lipids in leukemia. We analyzed leukemia samples from 12 patients with six distinct subtypes, alongside CD34<jats:sup>+<\/jats:sup>, B, T cells, monocytes and granulocytes from 3 healthy donors, by conducting high spatial resolution Raman imaging on 324 cells. We developed a single-cell phenotyping algorithm (incorporating cellular area, protein, nucleic acid, saturated and unsaturated lipid content) to distinguish leukemia subtypes. Finally, using H-SRPI and RNA-seq transcriptomics, we uncovered the critical role of lipid composition in leukemia cells across subtype classifications.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The single-cell phenotyping algorithm to distinguish leukemia subtypes, achieving 88.21% accuracy. H-SRPI and RNA-seq transcriptomes revealed elevated saturated and unsaturated lipid levels in acute myeloid leukemia (AML); AML-M3 favored lipid desaturation, whereas AML-M5 upregulated saturated lipid synthesis and elongation. ALL had weaker lipid metabolism characteristics than AML.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Our study establishes H-SRPI as a label-free tool for metabolic profiling, enabling precise leukemia subclassification and revealing lipid metabolic heterogeneity as a potential therapeutic target.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2025.1662281","type":"journal-article","created":{"date-parts":[[2025,10,1]],"date-time":"2025-10-01T04:18:02Z","timestamp":1759292282000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":3,"title":["High-content stimulated Raman pathology imaging and transcriptomics reveal leukemia subtype-specific lipid metabolic 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