a Overview of the hierarchical nature of an Acute Myeloid Leukemia hierarchy, with leukemic stem cells (LSC) at the apex, differentiating into progenitors, and subsequently, blasts. b FACS plot of the OCI-AML8227 hierarchy according to their CD34/CD38 surface marker expression levels. P1 are cells deemed live, P2 excludes doublets and Blasts, Progenitors and LSC are annotated according to CD34/CD38 expression. c scMS sample creation overview of booster channel samples and single cells; single-cell TMTpro samples were created with four Blast, five LSC and five Progenitor cells in each pool, labeled randomly using fourteen available TMTpro channels before pooling with a 200-cell equivalent of the 126-labeled booster sample. d Conceptual overview of our scMS experimental pipeline; single cells are sorted into 384-well plates containing 1ul of lysis buffer, then digested, TMT labeled and multiplexed. Resulting samples are analyzed with LC–MS via FAIMSPro gas-phase fractionation and Orbitrap detection.