{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T23:24:25Z","timestamp":1788823465789,"version":"build-2803163510"},"reference-count":46,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2025,7,28]],"date-time":"2025-07-28T00:00:00Z","timestamp":1753660800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2025,7,28]],"date-time":"2025-07-28T00:00:00Z","timestamp":1753660800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["32171390"],"award-info":[{"award-number":["32171390"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["32201154"],"award-info":[{"award-number":["32201154"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["T2350610283"],"award-info":[{"award-number":["T2350610283"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["32071318"],"award-info":[{"award-number":["32071318"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["2023B1515020104"],"award-info":[{"award-number":["2023B1515020104"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["2024A1515011244"],"award-info":[{"award-number":["2024A1515011244"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["2024A1515012508"],"award-info":[{"award-number":["2024A1515012508"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["2024A1515012522"],"award-info":[{"award-number":["2024A1515012522"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003453","name":"Natural Science Foundation of Guangdong Province","doi-asserted-by":"publisher","award":["JCYJ20220818101405011"],"award-info":[{"award-number":["JCYJ20220818101405011"]}],"id":[{"id":"10.13039\/501100003453","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Small Methods"],"published-print":{"date-parts":[[2026,2]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Macrophage polarization into inflammatory (M1) and repairing\/healing (M2) functional phenotypes are fundamental mechanisms in immune defensive responses, tissue repair, and disease control. Conventional phenotyping approaches based on molecular biomarkers are limited by destructive protocols, static endpoint analyses, and a disregard for the biomechanical attributes of cells. In this study, an integrated artificial intelligence (AI)\u2010atomic force microscopy (AFM) platform is introduced that enables label\u2010free, mechanophenotyping of macrophages at single\u2010cell resolution. Using nanoscale force mapping, morphological and nanomechanical profiles are captured details, such as Young's modulus, adhesion, and sphericity, across diverse macrophage activation states. These profiles are interpreted through a deep neural network (DNN) trained with pixel\u2010wise data enhancement and a meta\u2010confidence estimator for dynamic, robust classification. The system accurately distinguishes na\u00efve (M0), M1, and M2 functional phenotypes of human macrophages, even across donor heterogeneity, in the absence of conventional immunolabeling. The method reveals mixed macrophage polarization states and correlates cytoskeletal remodeling with mechanical biomarkers, establishing a direct link between cellular mechanics and immune function. This platform introduces a dynamic, non\u2010destructive strategy for immune monitoring, redefining cellular mechanics as a critical dimension in diagnostic and therapeutic contexts, and laying the groundwork for the emerging field of mechanoimmunology.<\/jats:p>","DOI":"10.1002\/smtd.202500953","type":"journal-article","created":{"date-parts":[[2025,7,29]],"date-time":"2025-07-29T04:25:35Z","timestamp":1753763135000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["AFM\u2010Based Deep Learning Decodes Human Macrophage Mechanophenotypes"],"prefix":"10.1002","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-4884-1515","authenticated-orcid":false,"given":"Jiaxin","family":"Chen","sequence":"first","affiliation":[{"name":"Laboratory of Inflammation and Vaccines Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences  Shenzhen Guangdong 518055 China"},{"name":"Laboratory of Immunology and Nanomedicine Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences  Shenzhen Guangdong 518055 China"},{"name":"China\u2010Italy Joint Laboratory of Pharmacobiotechnology for Medical Immunomodulation Shenzhen Institutes of Advanced Technology  Shenzhen Guangdong 518055 China"},{"name":"Division of Allergy &amp; Immunology Department of Biosciences &amp; Medical Biology Paris Lodron University of Salzburg  Salzburg 5020 Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Wu","sequence":"additional","affiliation":[{"name":"School of Management Science and Engineering Anhui University of Finance &amp; Economics  Bengbu Anhui 233030 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenjie","family":"Yang","sequence":"additional","affiliation":[{"name":"Laboratory of Inflammation and Vaccines Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences  Shenzhen Guangdong 518055 China"},{"name":"China\u2010Italy Joint Laboratory of Pharmacobiotechnology for Medical Immunomodulation Shenzhen Institutes of Advanced Technology  Shenzhen Guangdong 518055 China"},{"name":"Department of Environmental Biological and Pharmaceutical Sciences and Technologies (DiSTABiF) Universit\u00e0 Degli Studi Della Campania \u201cLuigi Vanvitelli\u201d  Caserta 81100 Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-2238-3202","authenticated-orcid":false,"given":"Haonan","family":"Li","sequence":"additional","affiliation":[{"name":"Laboratory of Inflammation and Vaccines Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences  Shenzhen Guangdong 518055 China"},{"name":"Laboratory of Immunology and Nanomedicine Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences  Shenzhen Guangdong 518055 China"},{"name":"China\u2010Italy Joint Laboratory of Pharmacobiotechnology for Medical Immunomodulation Shenzhen Institutes of Advanced Technology  Shenzhen Guangdong 518055 China"},{"name":"College of Life Science Hebei Normal University  Shijiazhuang Hebei 050016 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-8042-7209","authenticated-orcid":false,"given":"Qi","family":"Li","sequence":"additional","affiliation":[{"name":"Laboratory of Inflammation and Vaccines Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences  Shenzhen Guangdong 518055 China"},{"name":"Laboratory of Immunology and Nanomedicine Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences  Shenzhen Guangdong 518055 China"},{"name":"China\u2010Italy Joint Laboratory of Pharmacobiotechnology for Medical Immunomodulation Shenzhen Institutes of Advanced Technology  Shenzhen Guangdong 518055 China"},{"name":"Institut Catal\u00e0 de Nanoci\u00e8ncia I Nanotecnologia (ICN2) CSIC and The Barcelona Institute of Science and Technology (BIST)  Campus UAB Barcelona 08193 Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-5772-2743","authenticated-orcid":false,"given":"Su","family":"Li","sequence":"additional","affiliation":[{"name":"Laboratory of Inflammation and Vaccines Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences  Shenzhen Guangdong 518055 China"},{"name":"Laboratory of Immunology and Nanomedicine Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences  Shenzhen Guangdong 518055 China"},{"name":"China\u2010Italy Joint Laboratory of Pharmacobiotechnology for Medical Immunomodulation Shenzhen Institutes of Advanced Technology  Shenzhen Guangdong 518055 China"},{"name":"Division of Allergy &amp; Immunology Department of Biosciences &amp; Medical Biology Paris Lodron University of Salzburg  Salzburg 5020 Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6617-6752","authenticated-orcid":false,"given":"Yingnan","family":"Liu","sequence":"additional","affiliation":[{"name":"Laboratory of Inflammation and Vaccines Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences  Shenzhen Guangdong 518055 China"},{"name":"Laboratory of Immunology and Nanomedicine Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences  Shenzhen Guangdong 518055 China"},{"name":"China\u2010Italy Joint Laboratory of Pharmacobiotechnology for Medical Immunomodulation Shenzhen Institutes of Advanced Technology  Shenzhen Guangdong 518055 China"},{"name":"Division of Allergy &amp; Immunology Department of Biosciences &amp; Medical Biology Paris Lodron University of Salzburg  Salzburg 5020 Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fangfang","family":"Liu","sequence":"additional","affiliation":[{"name":"Laboratory of Inflammation and Vaccines Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences  Shenzhen Guangdong 518055 China"},{"name":"Laboratory of Immunology and Nanomedicine Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences  Shenzhen Guangdong 518055 China"},{"name":"China\u2010Italy Joint Laboratory of Pharmacobiotechnology for Medical Immunomodulation Shenzhen Institutes of Advanced Technology  Shenzhen Guangdong 518055 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yunping","family":"Xu","sequence":"additional","affiliation":[{"name":"Institution of Transfusion Medicine Shenzhen Blood Center  Shenzhen Guangdong 518000 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6386-9649","authenticated-orcid":false,"given":"Yan\u2010Zhong","family":"Chang","sequence":"additional","affiliation":[{"name":"College of Life Science Hebei Normal University  Shijiazhuang Hebei 050016 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5416-085X","authenticated-orcid":false,"given":"Martin","family":"Himly","sequence":"additional","affiliation":[{"name":"Division of Allergy &amp; Immunology Department of Biosciences &amp; Medical Biology Paris Lodron University of Salzburg  Salzburg 5020 Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paola","family":"Italiani","sequence":"additional","affiliation":[{"name":"Institute of Biochemistry and Cell Biology National Research Council (CNR)  Napoli 80131 Italy"},{"name":"China\u2010Italy Joint Laboratory of Pharmacobiotechnology for Medical Immunomodulation (SIAT, CNR) Stazione Zoologica Anton Dohrn (SZN)  Napoli 80121 Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Diana","family":"Boraschi","sequence":"additional","affiliation":[{"name":"Laboratory of Inflammation and Vaccines Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences  Shenzhen Guangdong 518055 China"},{"name":"China\u2010Italy Joint Laboratory of Pharmacobiotechnology for Medical Immunomodulation Shenzhen Institutes of Advanced Technology  Shenzhen Guangdong 518055 China"},{"name":"Institute of Biochemistry and Cell Biology National Research Council (CNR)  Napoli 80131 Italy"},{"name":"China\u2010Italy Joint Laboratory of Pharmacobiotechnology for Medical Immunomodulation (SIAT, CNR) Stazione Zoologica Anton Dohrn (SZN)  Napoli 80121 Italy"},{"name":"Shenzhen University of Advanced Technology  Shenzhen Guangdong 518107 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2488-570X","authenticated-orcid":false,"given":"Guofang","family":"Zhang","sequence":"additional","affiliation":[{"name":"Laboratory of Inflammation and Vaccines Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences  Shenzhen Guangdong 518055 China"},{"name":"Laboratory of Immunology and Nanomedicine Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences  Shenzhen Guangdong 518055 China"},{"name":"China\u2010Italy Joint Laboratory of Pharmacobiotechnology for Medical Immunomodulation Shenzhen Institutes of Advanced Technology  Shenzhen Guangdong 518055 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0404-4783","authenticated-orcid":false,"given":"Massimiliano","family":"Galluzzi","sequence":"additional","affiliation":[{"name":"Laboratory of Inflammation and Vaccines Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences  Shenzhen Guangdong 518055 China"},{"name":"China\u2010Italy Joint Laboratory of Pharmacobiotechnology for Medical Immunomodulation Shenzhen Institutes of Advanced Technology  Shenzhen Guangdong 518055 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7242-4687","authenticated-orcid":false,"given":"Yang","family":"Li","sequence":"additional","affiliation":[{"name":"Laboratory of Inflammation and Vaccines Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences  Shenzhen Guangdong 518055 China"},{"name":"Laboratory of Immunology and Nanomedicine Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences  Shenzhen Guangdong 518055 China"},{"name":"China\u2010Italy Joint Laboratory of Pharmacobiotechnology for Medical Immunomodulation Shenzhen Institutes of Advanced Technology  Shenzhen Guangdong 518055 China"},{"name":"Key Laboratory of Biomedical Imaging Science and System Chinese Academy of Sciences, and State Key Laboratory of Biomedical Imaging Science and System  Shenzhen 518055 China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,7,28]]},"reference":[{"key":"e_1_2_9_1_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2014.06.008"},{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.12703\/P6-13"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1615\/CritRevImmunol.v32.i6.10"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2014.00514"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2014.01.006"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-bioeng-062117-121224"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1063\/5.0087699"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0145813"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1111\/cpr.13440"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.chemrev.0c00617"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/nnano.2008.100"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/natrevmats.2017.8"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1152\/japplphysiol.00958.2007"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-11-0247"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1529\/biophysj.106.088831"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1007\/s13244-018-0639-9"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1093\/bib\/bbaa257"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.media.2017.07.005"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsnano.4c07729"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-39085-1"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1816459115"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.nanolett.1c00003"},{"key":"e_1_2_9_23_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsami.4c04330"},{"key":"e_1_2_9_24_1","doi-asserted-by":"publisher","DOI":"10.3389\/fbioe.2023.1259979"},{"key":"e_1_2_9_25_1","doi-asserted-by":"publisher","DOI":"10.1039\/D2BM01351G"},{"key":"e_1_2_9_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmbbm.2016.01.022"},{"key":"e_1_2_9_27_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10237-015-0706-9"},{"key":"e_1_2_9_28_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms25031860"},{"key":"e_1_2_9_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.imbio.2017.02.006"},{"key":"e_1_2_9_30_1","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20151570"},{"key":"e_1_2_9_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2016.02.015"},{"key":"e_1_2_9_32_1","doi-asserted-by":"publisher","DOI":"10.1002\/path.4133"},{"key":"e_1_2_9_33_1","doi-asserted-by":"publisher","DOI":"10.1038\/nri3834"},{"key":"e_1_2_9_34_1","doi-asserted-by":"publisher","DOI":"10.1038\/ni.3153"},{"key":"e_1_2_9_35_1","doi-asserted-by":"publisher","DOI":"10.1038\/ni1001-907"},{"key":"e_1_2_9_36_1","doi-asserted-by":"publisher","DOI":"10.1155\/2015\/816460"},{"key":"e_1_2_9_37_1","doi-asserted-by":"publisher","DOI":"10.1101\/cshperspect.a016303"},{"key":"e_1_2_9_38_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41422-021-00495-9"},{"key":"e_1_2_9_39_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2011.05.006"},{"key":"e_1_2_9_40_1","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20190418"},{"key":"e_1_2_9_41_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41565-021-00952-x"},{"key":"e_1_2_9_42_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.langmuir.8b01554"},{"key":"e_1_2_9_43_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-05902-1"},{"key":"e_1_2_9_44_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-3495(02)75620-8"},{"key":"e_1_2_9_45_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-05383-0"},{"key":"e_1_2_9_46_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41565-022-01177-2"}],"container-title":["Small Methods"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/smtd.202500953","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/smtd.202500953","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/smtd.202500953","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,11]],"date-time":"2026-02-11T15:42:58Z","timestamp":1770824578000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/smtd.202500953"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,28]]},"references-count":46,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2026,2]]}},"alternative-id":["10.1002\/smtd.202500953"],"URL":"https:\/\/doi.org\/10.1002\/smtd.202500953","archive":["Portico"],"relation":{},"ISSN":["2366-9608","2366-9608"],"issn-type":[{"value":"2366-9608","type":"print"},{"value":"2366-9608","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,28]]},"assertion":[{"value":"2025-05-18","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-07-28","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"2500953"}}