Abstract: Acute myeloid leukemia(AML) is a highly heterogeneous hematological malignancy, in which leukemia stem cells(LSCs) serve as the core drivers of disease relapse and therapeutic resistance. The precise identification of LSCs is a prerequisite for developing effective therapeutic strategies. Although conventional single-marker identification systems—primarily centered on the CD34⁺CD38⁻ phenotype and antigens such as CD33 and CD123—established the methodological foundation, they fail to comprehensively encompass the functional LSCs compartment. This limitation arises from the profound clonal heterogeneity of AML and significant phenotypic overlap with normal hematopoietic stem cells(HSCs). Driven by the application of high-throughput multi-omics technologies, LSCs identification strategies have transitioned toward the systemic elucidation of multidimensional co-expression networks and dynamic cellular states. Therefore, advancing from conventional flow cytometric markers, the applications of single-cell transcriptomics, mass cytometry, and integrated multi-omics analyses in the discovery of novel LSCs markers are explored. Furthermore, the differential expression profiles between LSCs and HSCs, regulatory mechanisms of these markers, combinatorial detection networks, and their clinical translational value are systematically summarized, aiming to provide a reference for the clinical translation of precision diagnostics and therapeutics in AML.