{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T04:56:03Z","timestamp":1789016163723,"version":"build-2803163510"},"reference-count":35,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2024,3,6]],"date-time":"2024-03-06T00:00:00Z","timestamp":1709683200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Horizon2020 ERC PoC","award":["101100664"],"award-info":[{"award-number":["101100664"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Bioengineering"],"abstract":"<jats:p>Myelodysplastic syndromes (MDSs) are a group of potentially deadly diseases that affect the morphology and function of neutrophils. Rapid diagnosis of MDS is crucial for the initiation of treatment that can vastly improve disease outcome. In this work, we present a new approach for detecting morphological differences between neutrophils isolated from blood samples of high-risk MDS patients and blood bank donors (BBDs). Using fluorescent flow cytometry, neutrophils were stained with 2\u2032,7\u2032-dichlorofluorescin diacetate (DCF), which reacts with reactive oxygen species (ROS), and Hoechst, which binds to DNA. We observed that BBDs possessed two cell clusters (designated H and L), whereas MDS patients possessed a single cluster (L). Later, we used FACS to sort the H and the L cells and used interferometric phase microscopy (IPM) to image the cells without utilizing cell staining. IPM images showed that H cells are characterized by low optical path delay (OPD) in the nucleus relative to the cytoplasm, especially in cell vesicles containing ROS, whereas L cells are characterized by low OPD in the cytoplasm relative to the nucleus and no ROS-containing vesicles. Moreover, L cells present a higher average OPD and dry mass compared to H cells. When examining neutrophils from MDS patients and BBDs by IPM during flow, we identified ~20% of cells as H cells in BBDs in contrast to ~4% in MDS patients. These results indicate that IPM can be utilized for the diagnosis of complex hematological pathologies such as MDS.<\/jats:p>","DOI":"10.3390\/bioengineering11030256","type":"journal-article","created":{"date-parts":[[2024,3,6]],"date-time":"2024-03-06T04:03:50Z","timestamp":1709697830000},"page":"256","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Analyzing Blood Cells of High-Risk Myelodysplastic Syndrome Patients Using Interferometric Phase Microscopy and Fluorescent Flow Cytometry"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8590-8995","authenticated-orcid":false,"given":"Itay","family":"Barnea","sequence":"first","affiliation":[{"name":"Department of Biomedical Engineering, Tel Aviv University, Tel Aviv 6997801, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lior","family":"Luria","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, Tel Aviv University, Tel Aviv 6997801, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arik","family":"Girsault","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, Tel Aviv University, Tel Aviv 6997801, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ofira","family":"Dabah","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, Tel Aviv University, Tel Aviv 6997801, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5836-8033","authenticated-orcid":false,"given":"Matan","family":"Dudaie","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, Tel Aviv University, Tel Aviv 6997801, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3244-8742","authenticated-orcid":false,"given":"Simcha K.","family":"Mirsky","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, Tel Aviv University, Tel Aviv 6997801, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Drorit","family":"Merkel","sequence":"additional","affiliation":[{"name":"MDS Center, Sheba Medical Center, Ramat Gan 5266202, Israel"},{"name":"Faculty of Medicine, Tel Aviv University, Tel Aviv 6997801, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natan T.","family":"Shaked","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, Tel Aviv University, Tel Aviv 6997801, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,3,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"872","DOI":"10.1001\/jama.2022.14578","article-title":"Diagnosis and Treatment of Myelodysplastic Syndromes: A Review","volume":"328","author":"Sekeres","year":"2022","journal-title":"JAMA"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2391","DOI":"10.1182\/blood-2016-03-643544","article-title":"The 2016 revision to the World Health Organization classification of myeloid neoplasms and acute leukemia","volume":"127","author":"Arber","year":"2016","journal-title":"Blood"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"3011","DOI":"10.1073\/pnas.1222861110","article-title":"Hematopoietic stem cell and progenitor cell mechanisms in myelodysplastic syndromes","volume":"110","author":"Pang","year":"2013","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1111\/j.1600-0609.2009.01299.x","article-title":"Platelet counts and haemorrhagic diathesis in patients with myelodysplastic syndromes","volume":"83","author":"Neukirchen","year":"2009","journal-title":"Eur. J. Haematol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"937","DOI":"10.1007\/s00277-016-2649-3","article-title":"Causes of death in 2877 patients with myelodysplastic syndromes","volume":"95","author":"Nachtkamp","year":"2016","journal-title":"Ann. Hematol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"471","DOI":"10.3109\/10428199409049637","article-title":"Neutrophils from patients with myelodysplastic syndromes: Relationship between impairment of granular contents, complement receptors, functional activities and disease status","volume":"13","author":"Moretti","year":"1994","journal-title":"Leuk. Lymphoma"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"538","DOI":"10.1111\/ijlh.13607","article-title":"How I investigate dysgranulopoiesis","volume":"43","author":"Shekhar","year":"2021","journal-title":"Int. J. Lab. Hematol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"752","DOI":"10.1016\/S0301-472X(03)00188-7","article-title":"Reduced expression of flavocytochrome b558, a component of the NADPH oxidase complex, in neutrophils from patients with myelodysplasia","volume":"31","author":"Fuhler","year":"2003","journal-title":"Exp. Hematol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"579","DOI":"10.1182\/blood-2004-01-0338","article-title":"A decision analysis of allogeneic bone marrow transplantation for the myelodysplastic syndromes: Delayed transplantation for low-risk myelodysplasia is associated with improved outcome","volume":"104","author":"Cutler","year":"2004","journal-title":"Blood"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"242","DOI":"10.1006\/excr.1994.1254","article-title":"Evaluation of oxidative processes in human pigment epithelial cells associated with retinal outer segment phagocytosis","volume":"214","author":"Miceli","year":"1994","journal-title":"Exp. Cell Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"240","DOI":"10.1016\/j.toxicon.2013.02.009","article-title":"Pro-inflammatory properties and neutrophil activation by Helicobacter pylori urease","volume":"69","author":"Uberti","year":"2013","journal-title":"Toxicon"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1385\/MB:19:2:125","article-title":"Using 2\u2032,7\u2032-dichlorodihydrofluorescein-diacetate to assess polysaccharides as immunomodulating agents","volume":"19","author":"Bland","year":"2001","journal-title":"Mol. Biotechnol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"121","DOI":"10.4103\/0970-1591.78430","article-title":"In vitro fertilization\/intracytoplasmic sperm injection for male infertility","volume":"27","author":"Merchant","year":"2011","journal-title":"Indian J. Urol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1757","DOI":"10.1364\/BOE.3.001757","article-title":"Generalized cell morphological parameters based on interferometric phase microscopy and their application to cell life cycle characterization","volume":"3","author":"Girshovitz","year":"2012","journal-title":"Biomed. Opt. Express"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"893","DOI":"10.1002\/cyto.a.23189","article-title":"Automated analysis of individual sperm cells using stain-free interferometric phase microscopy and machine learning","volume":"91","author":"Mirsky","year":"2017","journal-title":"Cytometry A"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1002\/cyto.a.24227","article-title":"Real-Time Stain-Free Classification of Cancer Cells and Blood Cells Using Interferometric Phase Microscopy and Machine Learning","volume":"99","author":"Nissim","year":"2021","journal-title":"Cytometry A"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2300433","DOI":"10.1002\/aisy.202300433","article-title":"Label-Free Imaging Flow Cytometry for Cell Classification Based on Multiple Interferometric Projections Using Deep Learning","volume":"6","author":"Cohen","year":"2024","journal-title":"Adv. Intell. Syst."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1800761","DOI":"10.1002\/advs.201800761","article-title":"Label-Free High-Throughput Leukemia Detection by Holographic Microscopy","volume":"5","author":"Ugele","year":"2018","journal-title":"Adv. Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"470","DOI":"10.1002\/cyto.a.24703","article-title":"Sperm-cell DNA fragmentation prediction using label-free quantitative phase imaging and deep learning","volume":"103","author":"Noy","year":"2023","journal-title":"Cytometry A"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"611679","DOI":"10.3389\/fphy.2020.611679","article-title":"Low-coherence shearing interferometry with constant off-axis angle","volume":"8","author":"Rongli","year":"2021","journal-title":"Front. Phys."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"8773","DOI":"10.1364\/OE.23.008773","article-title":"Fast phase processing in off-axis holography using multiplexing with complex encoding and live-cell fluctuation map calculation in real-time","volume":"23","author":"Girshovitz","year":"2015","journal-title":"Opt. Express"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Ban, B., Min, E., Ahn, Y., Popescu, G., and Jung, W. (2018). Effect of tissue staining in quantitative phase imaging. J. Biophotonics, 11.","DOI":"10.1002\/jbio.201700402"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/j.cll.2014.10.007","article-title":"White blood cell counts: Reference methodology","volume":"35","author":"George","year":"2015","journal-title":"Clin. Lab. Med."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1053\/j.seminhematol.2017.06.001","article-title":"Optimizing the use of hypomethylating agents in myelodysplastic syndromes: Selecting the candidate, predicting the response, and enhancing the activity","volume":"54","author":"Madanat","year":"2017","journal-title":"Semin. Hematol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1096","DOI":"10.1182\/blood-2018-10-844696","article-title":"Treatment of MDS","volume":"133","author":"Platzbecker","year":"2019","journal-title":"Blood"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1120","DOI":"10.1136\/jcp.36.10.1120","article-title":"Defective neutrophil function and microbicidal mechanisms in the myelodysplastic disorders","volume":"36","author":"Martin","year":"1983","journal-title":"J. Clin. Pathol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/S0165-4608(99)00188-0","article-title":"17p- syndrome arising from a novel dicentric translocation in a patient with acute myeloid leukemia","volume":"118","author":"Watson","year":"2000","journal-title":"Cancer Genet. Cytogenet."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"C1355","DOI":"10.1152\/ajpcell.00224.2002","article-title":"Glutamine delays spontaneous apoptosis in neutrophils","volume":"284","author":"Schumacher","year":"2003","journal-title":"Am. J. Physiol. Cell Physiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1083\/jcb.200806072","article-title":"Histone hypercitrullination mediates chromatin decondensation and neutrophil extracellular trap formation","volume":"184","author":"Wang","year":"2009","journal-title":"J. Cell Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1459","DOI":"10.3324\/haematol.2012.063420","article-title":"Infections in myelodysplastic syndromes","volume":"97","author":"Toma","year":"2012","journal-title":"Haematologica"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"577","DOI":"10.1038\/nrmicro1710","article-title":"Beneficial suicide: Why neutrophils die to make NETs","volume":"5","author":"Brinkmann","year":"2007","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"3007","DOI":"10.1182\/blood.V92.9.3007","article-title":"Inside the neutrophil phagosome: Oxidants, myeloperoxidase, and bacterial killing","volume":"92","author":"Hampton","year":"1998","journal-title":"Blood"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1532","DOI":"10.1126\/science.1092385","article-title":"Neutrophil extracellular traps kill bacteria","volume":"303","author":"Brinkmann","year":"2004","journal-title":"Science"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1083\/jcb.200606027","article-title":"Novel cell death program leads to neutrophil extracellular traps","volume":"176","author":"Fuchs","year":"2007","journal-title":"J. Cell Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1182\/bloodadvances.2021005721","article-title":"Impaired formation of neutrophil extracellular traps in patients with MDS","volume":"6","author":"Brings","year":"2022","journal-title":"Blood Adv."}],"container-title":["Bioengineering"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2306-5354\/11\/3\/256\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T14:09:48Z","timestamp":1760105388000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2306-5354\/11\/3\/256"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,3,6]]},"references-count":35,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2024,3]]}},"alternative-id":["bioengineering11030256"],"URL":"https:\/\/doi.org\/10.3390\/bioengineering11030256","relation":{},"ISSN":["2306-5354"],"issn-type":[{"value":"2306-5354","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,3,6]]}}}