Deoxygenation-free Digital Holographic Microscopy discriminates Sickle-Cell Disease in a pilot Colombian cohort via phase and shape metrics

Deoxygenation-free Digital Holographic Microscopy discriminates Sickle-Cell Disease in a pilot Colombian cohort via phase and shape metrics | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Deoxygenation-free Digital Holographic Microscopy discriminates Sickle-Cell Disease in a pilot Colombian cohort via phase and shape metrics Karina Ortega-Sanchez, Maria J. Gil-Herrera, Sayen Plaza, Alfonso Padilla-Vivanco, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7320264/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Rapid, reagent-free screening tools for sickle-cell disease (SCD) are urgently needed in low-resource regions where carrier frequency is high and laboratory infrastructure is scarce. We demonstrate digital holographic microscopy (DHM) as a label-free SCD tool in Colombia, imaging 885 individual erythrocytes (443 cells from venous blood of six SCD patients and 442 cells from 5 healthy donors). Two direct optical observables were extracted: phase difference (Δφ) relative to the surrounding medium and projected circularity index ( CI ), defined as the aspect ratio between the erythrocyte’s minor and major axes. Healthy cells are characterized by Δφ = 1.72 ± 0.30 (mean ± std) rad and CI = 0.94 ± 0.04, whereas SCD cells are distributed around Δφ = 3.00 ± 0.96 rad and CI = 0.70 ± 0.19. Despite some minimal overlap between both distributions, SCD cells are statistically distinguishable from healthy ones based on both Δφ and CI ( p 2.2 radians ∧ CI < 0.85) classified individual cells with 97.40% accuracy, 94.81% sensitivity, and 100% specificity. These results confirm DHM as a complementary approach to gold-standard methods, such as electrophoresis and HPLC, for SCD triage in resource-constrained settings. Biological sciences/Biological techniques Biological sciences/Biophysics Health sciences/Diseases Health sciences/Medical research Physical sciences/Optics and photonics Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7320264","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":508460758,"identity":"38061686-85bf-4c3d-bbc2-a6b642907fcf","order_by":0,"name":"Karina Ortega-Sanchez","email":"","orcid":"","institution":"EAFIT University","correspondingAuthor":false,"prefix":"","firstName":"Karina","middleName":"","lastName":"Ortega-Sanchez","suffix":""},{"id":508460759,"identity":"05812e0b-71f9-44b8-a461-3ff9de7f3df0","order_by":1,"name":"Maria J. Gil-Herrera","email":"","orcid":"","institution":"EAFIT University","correspondingAuthor":false,"prefix":"","firstName":"Maria","middleName":"J.","lastName":"Gil-Herrera","suffix":""},{"id":508460760,"identity":"e7eef85e-7e55-4d3c-bb72-3b8025a8b268","order_by":2,"name":"Sayen Plaza","email":"","orcid":"","institution":"Pontificia Universidad Católica de Valparaíso","correspondingAuthor":false,"prefix":"","firstName":"Sayen","middleName":"","lastName":"Plaza","suffix":""},{"id":508460761,"identity":"3703c304-d0e2-47fa-a708-166c3a5e6adc","order_by":3,"name":"Alfonso Padilla-Vivanco","email":"","orcid":"","institution":"Universidad Politécnica de Tulancingo, Tulancingo de Bravo","correspondingAuthor":false,"prefix":"","firstName":"Alfonso","middleName":"","lastName":"Padilla-Vivanco","suffix":""},{"id":508460762,"identity":"15e644f1-438a-49a7-8fa5-dfd52313edba","order_by":4,"name":"Earu Banoth","email":"","orcid":"","institution":"National Institute of Technology","correspondingAuthor":false,"prefix":"","firstName":"Earu","middleName":"","lastName":"Banoth","suffix":""},{"id":508460763,"identity":"b69d56cb-5aa1-4f2e-b794-fb52b7047a3f","order_by":5,"name":"Viviana Clavería","email":"","orcid":"","institution":"Pontificia Universidad Católica de Valparaíso","correspondingAuthor":false,"prefix":"","firstName":"Viviana","middleName":"","lastName":"Clavería","suffix":""},{"id":508460764,"identity":"518a95dd-09c5-49a3-845d-23a27ae85a60","order_by":6,"name":"Raúl Castañeda","email":"","orcid":"","institution":"EAFIT University","correspondingAuthor":false,"prefix":"","firstName":"Raúl","middleName":"","lastName":"Castañeda","suffix":""},{"id":508460765,"identity":"2c1e745f-caa8-4e0d-a272-b8a9c3a0f605","order_by":7,"name":"Ana Doblas","email":"","orcid":"","institution":"University of Massachussets Dartmouth","correspondingAuthor":false,"prefix":"","firstName":"Ana","middleName":"","lastName":"Doblas","suffix":""},{"id":508460766,"identity":"24bf824a-ed1c-4b64-a952-ef6d0b914e12","order_by":8,"name":"Carlos Trujillo","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA9UlEQVRIiWNgGAWjYNACHgkZPgbmAwwMNhIgrgEB5cxgLTxsDGwJDAxpEhJEagFqYmPgAapMYyCshb/9/LHPPDIWPGzsZz4++JFgUcfA3rxNgqHGBqcWiTPJzLN5QA7jyd1s2JMAdBjPsTIJhmNpOLUYMCQzM4O1MORuk+D9AdQikWMmwdhwGLcW/sdQLfxvnv/8A7JF/g0BLRIwWyRy2Jh5QFokePBrkbjx2JhxDljLM2NpmQQJyTaetGKLBDx+4e9PfMzwtqdOjp8/+eHHNwl1/Pzshzfe+IAnxMCAsQeJwwYiEvBrAIIfBFWMglEwCkbBSAYAm8c9MlC1wCUAAAAASUVORK5CYII=","orcid":"","institution":"EAFIT University","correspondingAuthor":true,"prefix":"","firstName":"Carlos","middleName":"","lastName":"Trujillo","suffix":""}],"badges":[],"createdAt":"2025-08-07 15:23:29","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7320264/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7320264/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":91503887,"identity":"d146dddd-a893-4ec6-9a42-b824c82fe80e","added_by":"auto","created_at":"2025-09-17 08:02:54","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1000322,"visible":true,"origin":"","legend":"","description":"","filename":"REVISEDSCADHMCharacterizationDEF.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7320264/v1_covered_3ec1c29c-a781-409a-8acf-e2427524c343.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Deoxygenation-free Digital Holographic Microscopy discriminates Sickle-Cell Disease in a pilot Colombian cohort via phase and shape metrics","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-7320264/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7320264/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eRapid, reagent-free screening tools for sickle-cell disease (SCD) are urgently needed in low-resource regions where carrier frequency is high and laboratory infrastructure is scarce. We demonstrate digital holographic microscopy (DHM) as a label-free SCD tool in Colombia, imaging 885 individual erythrocytes (443 cells from venous blood of six SCD patients and 442 cells from 5 healthy donors). Two direct optical observables were extracted: phase difference (Δφ) relative to the surrounding medium and projected circularity index (\u003cem\u003eCI\u003c/em\u003e), defined as the aspect ratio between the erythrocyte\u0026rsquo;s minor and major axes. Healthy cells are characterized by Δφ\u0026thinsp;=\u0026thinsp;1.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30 (mean\u0026thinsp;\u0026plusmn;\u0026thinsp;std) rad and \u003cem\u003eCI\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.94\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04, whereas SCD cells are distributed around Δφ\u0026thinsp;=\u0026thinsp;3.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.96 rad and \u003cem\u003eCI\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19. Despite some minimal overlap between both distributions, SCD cells are statistically distinguishable from healthy ones based on both Δφ and CI (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001). A simple rule (Δφ\u0026thinsp;\u0026gt;\u0026thinsp;2.2 radians \u0026and; \u003cem\u003eCI\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.85) classified individual cells with 97.40% accuracy, 94.81% sensitivity, and 100% specificity. These results confirm DHM as a complementary approach to gold-standard methods, such as electrophoresis and HPLC, for SCD triage in resource-constrained settings.\u003c/p\u003e","manuscriptTitle":"Deoxygenation-free Digital Holographic Microscopy discriminates Sickle-Cell Disease in a pilot Colombian cohort via phase and shape metrics","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-03 18:41:40","doi":"10.21203/rs.3.rs-7320264/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"075b8196-886a-4475-a839-197e90e49487","owner":[],"postedDate":"September 3rd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":53998725,"name":"Biological sciences/Biological techniques"},{"id":53998726,"name":"Biological sciences/Biophysics"},{"id":53998727,"name":"Health sciences/Diseases"},{"id":53998728,"name":"Health sciences/Medical research"},{"id":53998729,"name":"Physical sciences/Optics and photonics"}],"tags":[],"updatedAt":"2025-09-17T07:54:32+00:00","versionOfRecord":[],"versionCreatedAt":"2025-09-03 18:41:40","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7320264","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7320264","identity":"rs-7320264","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}