{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T11:18:50Z","timestamp":1788434330972,"version":"build-2803163510"},"reference-count":31,"publisher":"American Chemical Society (ACS)","issue":"34","license":[{"start":{"date-parts":[[2026,8,21]],"date-time":"2026-08-21T00:00:00Z","timestamp":1787270400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,8,21]],"date-time":"2026-08-21T00:00:00Z","timestamp":1787270400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,8,21]],"date-time":"2026-08-21T00:00:00Z","timestamp":1787270400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/100022963","name":"Key Research and Development Program of Zhejiang Province","doi-asserted-by":"publisher","award":["2025C02187"],"award-info":[{"award-number":["2025C02187"]}],"id":[{"id":"10.13039\/100022963","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004731","name":"Natural Science Foundation of Zhejiang Province","doi-asserted-by":"publisher","award":["LZ24H180001"],"award-info":[{"award-number":["LZ24H180001"]}],"id":[{"id":"10.13039\/501100004731","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100018684","name":"National Health Commission of the People's Republic of China","doi-asserted-by":"publisher","award":["2025YFC2427200"],"award-info":[{"award-number":["2025YFC2427200"]}],"id":[{"id":"10.13039\/501100018684","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/na","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["62571489"],"award-info":[{"award-number":["62571489"]}],"id":[{"id":"10.13039\/na","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/na","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82572363"],"award-info":[{"award-number":["82572363"]}],"id":[{"id":"10.13039\/na","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Invasive fungal infections are among the most time-critical clinical conditions, yet antifungal susceptibility testing (AFST) remains fundamentally constrained by culture-dependent, population-averaged workflows that require 48\u201372 h. Here, we introduce a culture-free and rapid AFST platform based on dynamic, single-fungus-resolved phenotypic profiling enabled by AI-powered lens-free holographic imaging. The platform integrates \u223c10 mm2 field-of-view on-chip lens-free holographic imaging with a low-cost, easy-to-operate microfluidic chip and a U-Net-based reconstruction and segmentation pipeline, enabling parallel, time-resolved profiling of hundreds of individual fungal cells and their morphodynamic behaviors. Dynamic fungal growth, hyphal elongation, and morphological transitions are quantified to generate multiparameter phenotypic fingerprints, which are interpreted by a machine-learning model for rapid susceptibility classification within 6 h. To demonstrate the clinical translational capability, we applied it directly to clinical urine, swab, and blood-culture specimens without conventional fungal cultivation and isolation and achieved 95.8% category agreement and 4.2% minor error with the clinical reference standard. By shifting susceptibility testing from delayed, population-averaged end points toward rapid, single-fungus-resolved dynamic phenotyping, this work provides a scalable, culture-free platform for rapid phenotypic AFST with potential applications in point-of-care and resource-limited settings.<\/jats:p>","DOI":"10.1021\/acs.analchem.6c04772","type":"journal-article","created":{"date-parts":[[2026,8,22]],"date-time":"2026-08-22T15:21:51Z","timestamp":1787412111000},"page":"25442-25453","source":"Crossref","is-referenced-by-count":0,"title":["AI-Enabled\nOn-Chip Holographic Imaging for Culture-Free,\nSingle-Fungus-Resolved Antifungal Susceptibility Testing"],"prefix":"10.1021","volume":"98","author":[{"given":"Jiahao","family":"Xu","sequence":"first","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310027"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-6791-9261","authenticated-orcid":true,"given":"Ruowang","family":"Dong","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310027"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2571-1242","authenticated-orcid":true,"given":"Yunrui","family":"Zhang","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310027"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ziqi","family":"Wang","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310027"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Shen","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310027"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Keting","family":"Xu","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310027"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying","family":"Fu","sequence":"additional","affiliation":[{"name":"Zhejiang University , School of Medicine, ,","place":["Hangzhou, China, 310016"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1581-4982","authenticated-orcid":true,"given":"Di","family":"Wang","sequence":"additional","affiliation":[{"name":"School of Intelligent Science and Technology, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences , ,","place":["Hangzhou, China, 310024"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Chen","sequence":"additional","affiliation":[{"name":"Zhejiang University , School of Medicine, ,","place":["Hangzhou, China, 310016"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4369-8563","authenticated-orcid":true,"given":"Fenni","family":"Zhang","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310027"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,21]]},"reference":[{"key":"2026090306462249300_cit1","doi-asserted-by":"publisher","first-page":"e428","DOI":"10.1016\/S1473-3099(23)00692-8","article-title":"Global\nincidence and mortality of severe fungal disease","volume":"24","author":"Denning","year":"2024","journal-title":"Lancet Infect. Dis."},{"key":"2026090306462249300_cit2","doi-asserted-by":"publisher","first-page":"e782","DOI":"10.1016\/S1473-3099(24)00429-8","article-title":"Antifungal susceptibility\ntesting: applicability of methods and strategies for improving access\nin resource-constrained settings","volume":"24","author":"Kwizera","year":"2024","journal-title":"Lancet Infect.\nDis."},{"key":"2026090306462249300_cit3","doi-asserted-by":"publisher","first-page":"720609","DOI":"10.3389\/fcimb.2021.720609","article-title":"Can We Improve Antifungal\nSusceptibility Testing?","volume":"11","author":"Durand","year":"2021","journal-title":"Front. Cell. Infect.\nMicrobiol."},{"key":"2026090306462249300_cit4","doi-asserted-by":"publisher","DOI":"10.1002\/smtd.202100713","article-title":"Paving the Way to Overcome\nAntifungal Drug Resistance: Current Practices and Novel Developments\nfor Rapid and Reliable Antifungal Susceptibility Testing","volume":"5","author":"Heuer","year":"2021","journal-title":"Small Methods"},{"key":"2026090306462249300_cit5","doi-asserted-by":"publisher","first-page":"3852","DOI":"10.1016\/j.apsb.2025.06.013","article-title":"Next-generation\nantifungal drugs: Mechanisms, efficacy, and clinical prospects","volume":"15","author":"Lu","year":"2025","journal-title":"Acta Pharm. Sin. 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Microbiol."},{"key":"2026090306462249300_cit8","doi-asserted-by":"publisher","first-page":"722","DOI":"10.3390\/jof9070722","article-title":"Development of an\nImaging Flow Cytometry\nMethod for Fungal Cytological Profiling and Its Potential Application\nin Antifungal Drug Development","volume":"9","author":"McMahon","year":"2023","journal-title":"J. Fungi"},{"key":"2026090306462249300_cit9","doi-asserted-by":"publisher","first-page":"2586","DOI":"10.1128\/JCM.43.6.2586-2589.2005","article-title":"Flow cytometry susceptibility testing\nfor the antifungal caspofungin","volume":"43","author":"Mitchell","year":"2005","journal-title":"J. Clin. Microbiol."},{"key":"2026090306462249300_cit10","doi-asserted-by":"publisher","first-page":"1305","DOI":"10.1039\/D3CC04446G","article-title":"Photonic Si microwell architectures\nfor rapid antifungal susceptibility determination of Candida auris","volume":"60","author":"Heuer","year":"2024","journal-title":"Chem. Commun."},{"key":"2026090306462249300_cit11","doi-asserted-by":"publisher","first-page":"2560","DOI":"10.1021\/acsinfecdis.0c00234","article-title":"Antifungal\nSusceptibility\nTesting of Aspergillus niger on Silicon Microwells by Intensity-Based\nReflectometric Interference Spectroscopy","volume":"6","author":"Heuer","year":"2020","journal-title":"ACS\nInfect. Dis."},{"key":"2026090306462249300_cit12","doi-asserted-by":"publisher","first-page":"2980","DOI":"10.1021\/acs.nanolett.3c03781","article-title":"An Optical\nFiber-Based Nanomotion Sensor for Rapid Antibiotic and Antifungal\nSusceptibility Tests","volume":"24","author":"Zhou","year":"2024","journal-title":"Nano Lett."},{"key":"2026090306462249300_cit13","doi-asserted-by":"publisher","first-page":"9099","DOI":"10.1038\/s41598-017-09329-4","article-title":"Potential Use of MALDI-ToF Mass Spectrometry\nfor Rapid Detection of Antifungal Resistance in the Human Pathogen\nCandida glabrata","volume":"7","author":"Vella","year":"2017","journal-title":"Sci. Rep"},{"key":"2026090306462249300_cit14","doi-asserted-by":"publisher","first-page":"15556","DOI":"10.1021\/acs.analchem.3c02243","article-title":"Rapid Antifungal Susceptibility\nTesting Based on Single-Cell Metabolism Analysis Using Stimulated\nRaman Scattering Imaging","volume":"95","author":"Chen","year":"2023","journal-title":"Anal. 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Sci."},{"key":"2026090306462249300_cit23","doi-asserted-by":"publisher","first-page":"3335","DOI":"10.1021\/acssensors.2c01277","article-title":"Multiplexed Chemical\nSensing CMOS Imager","volume":"7","author":"Wang","year":"2022","journal-title":"ACS\nSens."},{"key":"2026090306462249300_cit24","doi-asserted-by":"publisher","first-page":"14875","DOI":"10.1021\/acs.nanolett.4c04487","article-title":"Lensless On-Chip Chemiluminescence Imaging for High-Throughput\nSingle-Cell\nHeterogeneity Analysis","volume":"24","author":"Yang","year":"2024","journal-title":"Nano Lett."},{"key":"2026090306462249300_cit25","doi-asserted-by":"publisher","first-page":"71","DOI":"10.1021\/acssensors.2c01721","article-title":"Visual-Olfactory\nSynergistic Perception Based on Dual-Focus Imaging and a Bionic Learning\nArchitecture","volume":"8","author":"Cui","year":"2023","journal-title":"ACS Sens."},{"key":"2026090306462249300_cit26","doi-asserted-by":"publisher","first-page":"4872","DOI":"10.1109\/JSEN.2025.3645895","article-title":"Lensless CMOS-Coupled Platform for\nMiniaturized On-Chip\nElectrochemiluminescence Imaging and Sensing","volume":"26","author":"Fang","year":"2026","journal-title":"IEEE Sens. J."},{"key":"2026090306462249300_cit27","doi-asserted-by":"publisher","first-page":"10","DOI":"10.3390\/jof4010010","article-title":"Candida\nalbicans Hyphae: From Growth Initiation to Invasion","volume":"4","author":"Desai","year":"2018","journal-title":"J. Fungi"},{"key":"2026090306462249300_cit28","doi-asserted-by":"publisher","first-page":"317","DOI":"10.1016\/j.tim.2004.05.008","article-title":"The distinct\nmorphogenic states of Candida albicans","volume":"12","author":"Sudbery","year":"2004","journal-title":"Trends\nMicrobiol."},{"key":"2026090306462249300_cit29","doi-asserted-by":"publisher","first-page":"17","DOI":"10.3390\/jof5010017","article-title":"Linking Cellular\nMorphogenesis with Antifungal Treatment and Susceptibility in Candida\nPathogens","volume":"5","author":"Sharma","year":"2019","journal-title":"J. Fungi"},{"key":"2026090306462249300_cit30","doi-asserted-by":"publisher","first-page":"112","DOI":"10.1038\/nrmicro2711","article-title":"Candida albicans\nmorphogenesis and host defence: discriminating invasion from colonization","volume":"10","author":"Gow","year":"2012","journal-title":"Nat. Rev. Microbiol."},{"key":"2026090306462249300_cit31","doi-asserted-by":"publisher","first-page":"4294","DOI":"10.4161\/cc.22608","article-title":"Regulatory circuitry\ngoverning morphogenesis in Saccharomyces cerevisiae and Candida albicans","volume":"11","author":"Shapiro","year":"2012","journal-title":"Cell Cycle"}],"container-title":["Analytical Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.analchem.6c04772","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/ancham\/article-pdf\/98\/34\/25442\/66989903\/acs.analchem.6c04772.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/ancham\/article-pdf\/98\/34\/25442\/66989903\/acs.analchem.6c04772.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T10:46:27Z","timestamp":1788432387000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/ancham\/article\/98\/34\/25442\/5285958\/AI-Enabled-On-Chip-Holographic-Imaging-for-Culture"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,21]]},"references-count":31,"journal-issue":{"issue":"34","published-online":{"date-parts":[[2026,8,21]]},"published-print":{"date-parts":[[2026,9,1]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.analchem.6c04772","relation":{},"ISSN":["0003-2700","1520-6882"],"issn-type":[{"value":"0003-2700","type":"print"},{"value":"1520-6882","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,21]]}}}