{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T21:00:40Z","timestamp":1792184440171,"version":"4.4.0"},"reference-count":80,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2024,6,12]],"date-time":"2024-06-12T00:00:00Z","timestamp":1718150400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Danish Cancer Society"},{"name":"Max W\u00f8rzner and wife Inger W\u00f8rzner\u2019s Foundation"},{"name":"Department of Clinical Medicine, AU"},{"name":"Toyota Foundation, Dagmar Marshall\u2019s Foundation"},{"name":"Eva and Henry Fr\u00e6nkel\u2019s Memorial Foundation"},{"name":"Poul and Ellen Hertz\u2019s Foundation"},{"name":"Aase and Ejnar Danielsen\u2019s Foundation"},{"name":"Family Hede Nielsen Foundation"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Acute myeloid leukemia (AML) is a heterogenous blood cancer with a dismal prognosis. It emanates from leukemic stem cells (LSCs) arising from the genetic transformation of hematopoietic stem cells (HSCs). LSCs hold prognostic value, but their molecular and immunophenotypic heterogeneity poses challenges: there is no single marker for identifying all LSCs across AML samples. We hypothesized that imaging flow cytometry (IFC) paired with artificial intelligence-driven image analysis could visually distinguish LSCs from HSCs based solely on morphology. Initially, a seven-color IFC panel was employed to immunophenotypically identify LSCs and HSCs in bone marrow samples from five AML patients and ten healthy donors, respectively. Next, we developed convolutional neural network (CNN) models for HSC-LSC discrimination using brightfield (BF), side scatter (SSC), and DNA images. Classification using only BF images achieved 86.96% accuracy, indicating significant morphological differences. Accuracy increased to 93.42% when combining BF with DNA images, highlighting differences in nuclear morphology, although DNA images alone were inadequate for accurate HSC-LSC discrimination. Model development using SSC images revealed minor granularity differences. Performance metrics varied substantially between AML patients, indicating considerable morphologic variations among LSCs. Overall, we demonstrate proof-of-concept results for accurate CNN-based HSC-LSC differentiation, instigating the development of a novel technique within AML monitoring.<\/jats:p>","DOI":"10.3390\/ijms25126465","type":"journal-article","created":{"date-parts":[[2024,6,12]],"date-time":"2024-06-12T06:47:30Z","timestamp":1718174850000},"page":"6465","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Imaging Flow Cytometry and Convolutional Neural Network-Based Classification Enable Discrimination of Hematopoietic and Leukemic Stem Cells in Acute Myeloid Leukemia"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0166-8558","authenticated-orcid":false,"given":"Trine Engelbrecht","family":"Hybel","sequence":"first","affiliation":[{"name":"Department of Hematology, Aarhus University Hospital, 8200 Aarhus N, Denmark"},{"name":"Department of Clinical Medicine, Aarhus University, 8200 Aarhus N, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sofie Hesselberg","family":"Jensen","sequence":"additional","affiliation":[{"name":"Department of Hematology, Aarhus University Hospital, 8200 Aarhus N, Denmark"},{"name":"Department of Clinical Medicine, Aarhus University, 8200 Aarhus N, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew A.","family":"Rodrigues","sequence":"additional","affiliation":[{"name":"Amnis Flow Cytometry, Cytek Biosciences, Seattle, WA 98119, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas Engelbrecht","family":"Hybel","sequence":"additional","affiliation":[{"name":"Department of Hematology, Aarhus University Hospital, 8200 Aarhus N, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maya Nautrup","family":"Pedersen","sequence":"additional","affiliation":[{"name":"Department of Hematology, Aarhus University Hospital, 8200 Aarhus N, Denmark"},{"name":"Department of Clinical Medicine, Aarhus University, 8200 Aarhus N, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Signe H\u00e5kansson","family":"Qvick","sequence":"additional","affiliation":[{"name":"Department of Hematology, Aarhus University Hospital, 8200 Aarhus N, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marie Hairing","family":"Enemark","sequence":"additional","affiliation":[{"name":"Department of Hematology, Aarhus University Hospital, 8200 Aarhus N, Denmark"},{"name":"Department of Clinical Medicine, Aarhus University, 8200 Aarhus N, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marie","family":"Bill","sequence":"additional","affiliation":[{"name":"Department of Hematology, Aarhus University Hospital, 8200 Aarhus N, Denmark"},{"name":"Department of Clinical Medicine, Aarhus University, 8200 Aarhus N, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9609-8991","authenticated-orcid":false,"given":"Carina Agerbo","family":"Rosenberg","sequence":"additional","affiliation":[{"name":"Department of Hematology, Aarhus University Hospital, 8200 Aarhus N, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5089-3271","authenticated-orcid":false,"given":"Maja","family":"Ludvigsen","sequence":"additional","affiliation":[{"name":"Department of Hematology, Aarhus University Hospital, 8200 Aarhus N, Denmark"},{"name":"Department of Clinical Medicine, Aarhus University, 8200 Aarhus N, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,6,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e2300229","DOI":"10.1200\/GO.23.00229","article-title":"Burden of AML, 1990\u20132019: Estimates From the Global Burden of Disease Study","volume":"9","author":"Jani","year":"2023","journal-title":"JCO Glob. Oncol."},{"key":"ref_2","first-page":"1454","article-title":"Escape from Treatment; the Different Faces of Leukemic Stem Cells and Therapy Resistance in Acute Myeloid Leukemia","volume":"11","author":"Denkers","year":"2021","journal-title":"Front. Oncol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1315","DOI":"10.1038\/nbt1350","article-title":"Chemotherapy-Resistant Human AML Stem Cells Home to and Engraft within the Bone-Marrow Endosteal Region","volume":"25","author":"Ishikawa","year":"2007","journal-title":"Nat. Biotechnol."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Terwijn, M., Zeijlemaker, W., Kelder, A., Rutten, A.P., Snel, A.N., Scholten, W.J., Pabst, T., Verhoef, G., L\u00f6wenberg, B., and Zweegman, S. (2014). Leukemic Stem Cell Frequency: A Strong Biomarker for Clinical Outcome in Acute Myeloid Leukemia. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0107587"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1211","DOI":"10.1007\/s12015-021-10308-6","article-title":"Acute Myeloid Leukemia Stem Cells: Origin, Characteristics, and Clinical Implications","volume":"18","author":"Long","year":"2022","journal-title":"Stem Cell Rev. Rep."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e441","DOI":"10.1038\/bcj.2016.50","article-title":"Acute Myeloid Leukemia: A Comprehensive Review and 2016 Update","volume":"6","year":"2016","journal-title":"Blood Cancer J."},{"key":"ref_7","first-page":"38","article-title":"Acute Myeloid Leukemia: A Review","volume":"103","author":"Pelcovits","year":"2020","journal-title":"R. I. Med. J."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"n2026","DOI":"10.1136\/bmj.n2026","article-title":"Advances in Acute Myeloid Leukemia","volume":"375","author":"Newell","year":"2021","journal-title":"BMJ"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"502","DOI":"10.1002\/ajh.26822","article-title":"Acute Myeloid Leukemia: 2023 Update on Diagnosis, Risk-Stratification, and Management","volume":"98","author":"Shimony","year":"2023","journal-title":"Am. J. Hematol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"424","DOI":"10.1182\/blood-2016-08-733196","article-title":"Diagnosis and Management of AML in Adults: 2017 ELN Recommendations from an International Expert Panel","volume":"129","author":"Estey","year":"2017","journal-title":"Blood"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1182\/blood-2009-07-235358","article-title":"Diagnosis and Management of Acute Myeloid Leukemia in Adults: Recommendations from an International Expert Panel, on Behalf of the European LeukemiaNet","volume":"115","author":"Estey","year":"2010","journal-title":"Blood"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1182\/blood-2015-08-604520","article-title":"An Update of Current Treatments for Adult Acute Myeloid Leukemia","volume":"127","author":"Dombret","year":"2016","journal-title":"Blood"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1136","DOI":"10.1056\/NEJMra1406184","article-title":"Acute Myeloid Leukemia","volume":"373","author":"Weisdorf","year":"2015","journal-title":"N. Engl. J. Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"988","DOI":"10.1038\/leu.2014.327","article-title":"Prognostic Value of Monitoring a Candidate Immunophenotypic Leukaemic Stem\/Progenitor Cell Population in Patients Allografted for Acute Myeloid Leukaemia","volume":"29","author":"Bradbury","year":"2015","journal-title":"Leukemia"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1700","DOI":"10.1038\/sj.leu.2404754","article-title":"Aberrant Marker Expression Patterns on the CD34+CD38- Stem Cell Compartment in Acute Myeloid Leukemia Allows to Distinguish the Malignant from the Normal Stem Cell Compartment Both at Diagnosis and in Remission","volume":"21","author":"Moshaver","year":"2007","journal-title":"Leukemia"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1345","DOI":"10.1182\/blood.2022016867","article-title":"Diagnosis and Management of AML in Adults: 2022 Recommendations from an International Expert Panel on Behalf of the ELN","volume":"140","author":"Wei","year":"2022","journal-title":"Blood"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"3345","DOI":"10.1182\/blood-2014-05-577593","article-title":"Defining Minimal Residual Disease in Acute Myeloid Leukemia: Which Platforms Are Ready for \u201cPrime Time\u201d?","volume":"124","author":"Grimwade","year":"2014","journal-title":"Blood"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1275","DOI":"10.1182\/blood-2017-09-801498","article-title":"Minimal\/Measurable Residual Disease in AML: A Consensus Document from the European LeukemiaNet MRD Working Party","volume":"131","author":"Schuurhuis","year":"2018","journal-title":"Blood"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2810","DOI":"10.3324\/haematol.2022.282034","article-title":"The Present and Future of Measurable Residual Disease Testing in Acute Myeloid Leukemia","volume":"107","author":"Blachly","year":"2022","journal-title":"Haematologica"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1158\/1078-0432.CCR-22-2237","article-title":"Prognostic Impact of Unsupervised Early Assessment of Bulk and Leukemic Stem Cell Measurable Residual Disease in Acute Myeloid Leukemia","volume":"29","author":"Canali","year":"2023","journal-title":"Clin. Cancer Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1102","DOI":"10.1038\/s41375-018-0326-3","article-title":"CD34(+)CD38(\u2212) Leukemic Stem Cell Frequency to Predict Outcome in Acute Myeloid Leukemia","volume":"33","author":"Zeijlemaker","year":"2019","journal-title":"Leukemia"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"e66","DOI":"10.1002\/cpcy.66","article-title":"Immunophenotypic Detection of Measurable Residual (Stem Cell) Disease Using LAIP Approach in Acute Myeloid Leukemia","volume":"91","author":"Zeijlemaker","year":"2019","journal-title":"Curr. Protoc. Cytom."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1777","DOI":"10.1038\/sj.leu.2401903","article-title":"The Interleukin-3 Receptor Alpha Chain Is a Unique Marker for Human Acute Myelogenous Leukemia Stem Cells","volume":"14","author":"Jordan","year":"2000","journal-title":"Leukemia"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1038\/s41375-018-0180-3","article-title":"Coexpression Profile of Leukemic Stem Cell Markers for Combinatorial Targeted Therapy in AML","volume":"33","author":"Haubner","year":"2019","journal-title":"Leukemia"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2659","DOI":"10.1182\/blood-2007-03-083048","article-title":"The Novel AML Stem Cell Associated Antigen CLL-1 Aids in Discrimination between Normal and Leukemic Stem Cells","volume":"110","author":"Kelder","year":"2007","journal-title":"Blood"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"645","DOI":"10.1038\/367645a0","article-title":"A Cell Initiating Human Acute Myeloid Leukaemia after Transplantation into SCID Mice","volume":"367","author":"Lapidot","year":"1994","journal-title":"Nature"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"730","DOI":"10.1038\/nm0797-730","article-title":"Human Acute Myeloid Leukemia Is Organized as a Hierarchy That Originates from a Primitive Hematopoietic Cell","volume":"3","author":"Bonnet","year":"1997","journal-title":"Nat. Med."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1577","DOI":"10.1182\/blood-2016-10-696054","article-title":"Biology and Relevance of Human Acute Myeloid Leukemia Stem Cells","volume":"129","author":"Thomas","year":"2017","journal-title":"Blood"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"452","DOI":"10.1111\/bjh.18094","article-title":"Immunophenotypically Defined Stem Cell Subsets in Paediatric AML Are Highly Heterogeneous and Demonstrate Differences in BCL-2 Expression by Cytogenetic Subgroups","volume":"197","author":"Petersen","year":"2022","journal-title":"Br. J. Haematol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"382","DOI":"10.1002\/cyto.a.24284","article-title":"OMIP 072: A 15-Color Panel for Immunophenotypic Identification, Quantification, and Characterization of Leukemic Stem Cells in Children with Acute Myeloid Leukemia","volume":"99","author":"Petersen","year":"2021","journal-title":"Cytom. Part A J. Int. Soc. Anal. Cytol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"769","DOI":"10.1111\/bjh.15711","article-title":"Revisiting CLEC12A as Leukaemic Stem Cell Marker in AML: Highlighting the Necessity of Precision Diagnostics in Patients Eligible for Targeted Therapy","volume":"184","author":"Bill","year":"2019","journal-title":"Br. J. Haematol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2311","DOI":"10.1111\/jcmm.13519","article-title":"Mapping the CLEC12A Expression on Myeloid Progenitors in Normal Bone Marrow; Implications for Understanding CLEC12A-Related Cancer Stem Cell Biology","volume":"22","author":"Bill","year":"2018","journal-title":"J. Cell. Mol. Med."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Arnone, M., Konantz, M., Hanns, P., Paczulla Stanger, A.M., Bertels, S., Godavarthy, P.S., Christopeit, M., and Lengerke, C. (2020). Acute Myeloid Leukemia Stem Cells: The Challenges of Phenotypic Heterogeneity. Cancers, 12.","DOI":"10.3390\/cancers12123742"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"4639","DOI":"10.1039\/C6LC01063F","article-title":"Review: Imaging Technologies for Flow Cytometry","volume":"16","author":"Han","year":"2016","journal-title":"Lab Chip"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"649","DOI":"10.1016\/j.tibtech.2017.12.008","article-title":"Diagnostic Potential of Imaging Flow Cytometry","volume":"36","author":"Doan","year":"2018","journal-title":"Trends Biotechnol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1007\/978-1-4939-3302-0_2","article-title":"Principles of Amnis Imaging Flow Cytometry","volume":"1389","author":"Basiji","year":"2016","journal-title":"Methods Mol. Biol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"836","DOI":"10.1002\/cyto.a.23794","article-title":"Label-Free Identification of White Blood Cells Using Machine Learning","volume":"95","author":"Nassar","year":"2019","journal-title":"Cytom. Part A"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"414","DOI":"10.1038\/s41563-019-0339-y","article-title":"Leveraging Machine Vision in Cell-Based Diagnostics to Do More with Less","volume":"18","author":"Doan","year":"2019","journal-title":"Nat. Mater."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1002\/cyto.a.23987","article-title":"Label-Free Leukemia Monitoring by Computer Vision","volume":"97","author":"Doan","year":"2020","journal-title":"Cytom. Part A"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Shapiro, H.M. (2003). Practical Flow Cytometry, John Wiley & Sons.","DOI":"10.1002\/0471722731"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1177\/1947601911408076","article-title":"Genetic Abnormalities and Challenges in the Treatment of Acute Myeloid Leukemia","volume":"2","author":"Kumar","year":"2011","journal-title":"Genes Cancer"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1029","DOI":"10.1634\/stemcells.2006-0567","article-title":"Side Population Analysis Using a Violet-Excited Cell-Permeable DNA Binding Dye","volume":"25","author":"Telford","year":"2007","journal-title":"Stem Cells"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.leukres.2019.04.004","article-title":"Relationship between CD34\/CD38 and Side Population (SP) Defined Leukemia Stem Cell Compartments in Acute Myeloid Leukemia","volume":"81","author":"Moshaver","year":"2019","journal-title":"Leuk. Res."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"8443","DOI":"10.1158\/0008-5472.CAN-04-1659","article-title":"C-Type Lectin-like Molecule-1: A Novel Myeloid Cell Surface Marker Associated with Acute Myeloid Leukemia","volume":"64","author":"Bakker","year":"2004","journal-title":"Cancer Res."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1007\/978-1-4939-9650-6_15","article-title":"Immunophenotyping of Acute Myeloid Leukemia","volume":"2032","author":"Galera","year":"2019","journal-title":"Methods Mol. Biol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"538","DOI":"10.1038\/s42256-019-0101-9","article-title":"Human-Level Recognition of Blast Cells in Acute Myeloid Leukaemia with Convolutional Neural Networks","volume":"1","author":"Matek","year":"2019","journal-title":"Nat. Mach. Intell."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"951","DOI":"10.1172\/JCI113409","article-title":"Antigen CD34+ Marrow Cells Engraft Lethally Irradiated Baboons","volume":"81","author":"Berenson","year":"1988","journal-title":"J. Clin. Investig."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1603","DOI":"10.1038\/sj.leu.2402620","article-title":"CD34+ or CD34\u2212: Which Is the More Primitive?","volume":"16","author":"Engelhardt","year":"2002","journal-title":"Leukemia"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"3306","DOI":"10.1182\/blood.V88.9.3306.bloodjournal8893306","article-title":"Extended Long-Term Culture Reveals a Highly Quiescent and Primitive Human Hematopoietic Progenitor Population","volume":"88","author":"Hao","year":"1996","journal-title":"Blood"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1515","DOI":"10.1182\/blood.V83.6.1515.1515","article-title":"Lymphoid and Myeloid Differentiation of Single Human CD34+, HLA-DR+, CD38- Hematopoietic Stem Cells","volume":"83","author":"Huang","year":"1994","journal-title":"Blood"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1182\/blood.V93.1.96","article-title":"Single Adult Human CD34(+)\/Lin-\/CD38(-) Progenitors Give Rise to Natural Killer Cells, B-Lineage Cells, Dendritic Cells, and Myeloid Cells","volume":"93","author":"Miller","year":"1999","journal-title":"Blood"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1470","DOI":"10.1073\/pnas.93.4.1470","article-title":"Self-Renewal of Primitive Human Hematopoietic Cells (Long-Term-Culture-Initiating Cells) in Vitro and Their Expansion in Defined Medium","volume":"93","author":"Petzer","year":"1996","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1218","DOI":"10.1182\/blood.V77.6.1218.1218","article-title":"Sequential Generations of Hematopoietic Colonies Derived from Single Nonlineage-Committed CD34+CD38- Progenitor Cells","volume":"77","author":"Terstappen","year":"1991","journal-title":"Blood"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"640","DOI":"10.1002\/wsbm.86","article-title":"Hematopoietic Stem Cell: Self-Renewal versus Differentiation","volume":"2","author":"Seita","year":"2010","journal-title":"Wiley Interdiscip. Rev. Syst. Biol. Med."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"891","DOI":"10.1111\/bjh.16594","article-title":"Applicability and Reproducibility of Acute Myeloid Leukaemia Stem Cell Assessment in a Multi-Centre Setting","volume":"190","author":"Hanekamp","year":"2020","journal-title":"Br. J. Haematol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1038\/nature20598","article-title":"A 17-Gene Stemness Score for Rapid Determination of Risk in Acute Leukaemia","volume":"540","author":"Ng","year":"2016","journal-title":"Nature"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"1086","DOI":"10.1038\/nm.2415","article-title":"Stem Cell Gene Expression Programs Influence Clinical Outcome in Human Leukemia","volume":"17","author":"Eppert","year":"2011","journal-title":"Nat. Med."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1976","DOI":"10.1182\/blood-2009-02-206565","article-title":"Leukemia-Initiating Cells from Some Acute Myeloid Leukemia Patients with Mutated Nucleophosmin Reside in the CD34(-) Fraction","volume":"115","author":"Taussig","year":"2010","journal-title":"Blood"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"384","DOI":"10.1172\/JCI41495","article-title":"Human Acute Myelogenous Leukemia Stem Cells Are Rare and Heterogeneous When Assayed in NOD\/SCID\/IL2R\u03b3c-Deficient Mice","volume":"121","author":"Sarry","year":"2011","journal-title":"J. Clin. Investig."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1016\/j.ccr.2010.12.012","article-title":"Coexistence of LMPP-like and GMP-like Leukemia Stem Cells in Acute Myeloid Leukemia","volume":"19","author":"Goardon","year":"2011","journal-title":"Cancer Cell"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"1513","DOI":"10.1084\/jem.20151775","article-title":"Genetically Distinct Leukemic Stem Cells in Human CD34- Acute Myeloid Leukemia Are Arrested at a Hemopoietic Precursor-like Stage","volume":"213","author":"Quek","year":"2016","journal-title":"J. Exp. Med."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"1671","DOI":"10.1182\/blood-2016-02-695312","article-title":"Evolution of Acute Myelogenous Leukemia Stem Cell Properties after Treatment and Progression","volume":"128","author":"Ho","year":"2016","journal-title":"Blood"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"439","DOI":"10.1038\/leu.2015.252","article-title":"A Simple One-Tube Assay for Immunophenotypical Quantification of Leukemic Stem Cells in Acute Myeloid Leukemia","volume":"30","author":"Zeijlemaker","year":"2016","journal-title":"Leukemia"},{"key":"ref_64","doi-asserted-by":"crossref","unstructured":"Salek, M., Li, N., Chou, H.-P., Saini, K., Jovic, A., Jacobs, K.B., Johnson, C., Lu, V., Lee, E.J., and Chang, C. (2023). COSMOS: A Platform for Real-Time Morphology-Based, Label-Free Cell Sorting Using Deep Learning. Commun. Biol., 6.","DOI":"10.1038\/s42003-023-05325-9"},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"1246","DOI":"10.1126\/science.aan0096","article-title":"Ghost Cytometry","volume":"360","author":"Ota","year":"2018","journal-title":"Science"},{"key":"ref_66","doi-asserted-by":"crossref","unstructured":"Ng, D.P., Simonson, P.D., Tarnok, A., Lucas, F., Kern, W., Rolf, N., Bogdanoski, G., Green, C., Brinkman, R.R., and Czechowska, K. (2024). Recommendations for Using Artificial Intelligence in Clinical Flow Cytometry. Cytom. Part B Clin. Cytom., in press.","DOI":"10.1002\/cyto.b.22166"},{"key":"ref_67","doi-asserted-by":"crossref","unstructured":"James, G., Witten, D., Hastie, T., and Tibshirani, R. (2021). An Introduction to Statistical Learning, Springer.","DOI":"10.1007\/978-1-0716-1418-1"},{"key":"ref_68","doi-asserted-by":"crossref","unstructured":"Rodrigues, M.A., Probst, C.E., Zayats, A., Davidson, B., Riedel, M., Li, Y., and Venkatachalam, V. (2021). The in Vitro Micronucleus Assay Using Imaging Flow Cytometry and Deep Learning. NPJ Syst. Biol. Appl., 7.","DOI":"10.1038\/s41540-021-00179-5"},{"key":"ref_69","doi-asserted-by":"crossref","unstructured":"Stirling, D.R., Swain-Bowden, M.J., Lucas, A.M., Carpenter, A.E., Cimini, B.A., and Goodman, A. (2021). CellProfiler 4: Improvements in Speed, Utility and Usability. BMC Bioinform., 22.","DOI":"10.1186\/s12859-021-04344-9"},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1016\/j.ymeth.2016.08.018","article-title":"An Open-Source Solution for Advanced Imaging Flow Cytometry Data Analysis Using Machine Learning","volume":"112","author":"Hennig","year":"2017","journal-title":"Methods"},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"463","DOI":"10.1038\/s41467-017-00623-3","article-title":"Reconstructing Cell Cycle and Disease Progression Using Deep Learning","volume":"8","author":"Eulenberg","year":"2017","journal-title":"Nat. Commun."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"3572","DOI":"10.1038\/s41596-021-00549-7","article-title":"Deepometry, a Framework for Applying Supervised and Weakly Supervised Deep Learning to Imaging Cytometry","volume":"16","author":"Doan","year":"2021","journal-title":"Nat. Protoc."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1016\/j.ymeth.2016.10.002","article-title":"Imaging Flow Cytometry in the Assessment of Leukocyte-Platelet Aggregates","volume":"112","author":"Hui","year":"2017","journal-title":"Methods"},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"21381","DOI":"10.1073\/pnas.2001227117","article-title":"Objective Assessment of Stored Blood Quality by Deep Learning","volume":"117","author":"Doan","year":"2020","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"554","DOI":"10.1002\/cyto.b.21975","article-title":"Exploring Dyserythropoiesis in Patients with Myelodyspla Stic Syndrome by Imaging Flow Cytometry and Machine-Learning Assisted Morphometrics","volume":"100","author":"Rosenberg","year":"2020","journal-title":"Clin. Cytom. Part B"},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"9349","DOI":"10.1038\/s41598-024-59875-x","article-title":"Comparative Analysis of Feature-Based ML and CNN for Binucleated Erythroblast Quantification in Myelodysplastic Syndrome Patients Using Imaging Flow Cytometry Data","volume":"14","author":"Rosenberg","year":"2024","journal-title":"Sci. Rep."},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"1001","DOI":"10.1002\/cyto.a.22995","article-title":"Elimination of Erroneous Results in Flow Cytometry Caused by Antibody Binding to Fc Receptors on Human Monocytes and Macrophages","volume":"89","author":"Andersen","year":"2016","journal-title":"Cytom. Part A J. Int. Soc. Anal. Cytol."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"2996","DOI":"10.1016\/j.xphs.2020.07.008","article-title":"Advanced Characterization of Silicone Oil Droplets in Protein Therapeutics Using Artificial Intelligence Analysis of Imaging Flow Cytometry Data","volume":"109","author":"Probst","year":"2020","journal-title":"J. Pharm. Sci."},{"key":"ref_79","unstructured":"Abadi, M., Agarwal, A., Barham, P., Brevdo, E., Chen, Z., Citro, C., Corrado, G.S., Davis, A., Dean, J., and Devin, M. (2016). Tensorflow: Large-Scale Machine Learning on Heterogeneous Distributed Systems. arXiv."},{"key":"ref_80","unstructured":"Simonyan, K., and Zisserman, A. (2014). Very Deep Convolutional Networks for Large-Scale Image Recognition. arXiv."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/12\/6465\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T14:57:15Z","timestamp":1760108235000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/12\/6465"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,6,12]]},"references-count":80,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2024,6]]}},"alternative-id":["ijms25126465"],"URL":"https:\/\/doi.org\/10.3390\/ijms25126465","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,6,12]]}}}