Selective Lysosomal H2O2 – ROS Imaging with a Naphthalimide Probe Forming Hydroxylamine without Overoxidation

Selective Lysosomal H2O2 – ROS Imaging with a Naphthalimide Probe Forming Hydroxylamine without Overoxidation | 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 Research Article Selective Lysosomal H 2 O 2 – ROS Imaging with a Naphthalimide Probe Forming Hydroxylamine without Overoxidation Ricardo Flores-Cruz, Nitzya Ruiz-Robledo, Adriana Romo-Pérez, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8253737/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 Hydrogen peroxide (H 2 O 2 ) plays a central role in oxidative stress, signaling, and pathophysiology, yet its highly dynamic and compartmentalized distribution in living cells makes precise monitoring a major analytical challenge. Here, we present LysoH 2 O 2 , a naphthalimide-derived fluorescent probe specifically designed for lysosomal H 2 O 2 detection and imaging. The probe exploits a selective transformation of an amine group into a hydroxylamine moiety upon reaction with H 2 O 2 , leading to a pronounced fluorescence enhancement. This molecular design integrates a lysosome-targeting unit to ensure subcellular specificity, enabling real-time visualization of H 2 O 2 fluctuations in the lysosomal microenvironment. Confocal fluorescence microscopy demonstrates the ability of LysoH 2 O 2 to track endogenous and exogenous H 2 O 2 with high sensitivity and minimal cytotoxicity in living cells. Distinct from conventional boronate-based probes, LysoH 2 O 2 employs a novel, biocompatible chemical transformation to achieve sensitive and selective monitoring of H 2 O 2 within lysosomes. Cell dynamics Lipid Droplets Fluorescent Probes Fluorophore-Targeting Endoplasmic Reticulum Full Text Additional Declarations No competing interests reported. Supplementary Files SIfile.docx 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-8253737","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":560082554,"identity":"ad9a1374-659b-405e-b9fc-78de28d2b2d7","order_by":0,"name":"Ricardo Flores-Cruz","email":"","orcid":"","institution":"National Autonomous University of Mexico","correspondingAuthor":false,"prefix":"","firstName":"Ricardo","middleName":"","lastName":"Flores-Cruz","suffix":""},{"id":560082555,"identity":"6ff174bb-cd3e-40c1-be65-b2613d04f2b5","order_by":1,"name":"Nitzya Ruiz-Robledo","email":"","orcid":"","institution":"National Autonomous University of 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Overoxidation\u003c/p\u003e","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":"Cell dynamics, Lipid Droplets, Fluorescent Probes, Fluorophore-Targeting, Endoplasmic 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Confocal fluorescence microscopy demonstrates the ability of \u003cb\u003eLysoH\u003c/b\u003e\u003csub\u003e\u003cb\u003e2\u003c/b\u003e\u003c/sub\u003e\u003cb\u003eO\u003c/b\u003e\u003csub\u003e\u003cb\u003e2\u003c/b\u003e\u003c/sub\u003e to track endogenous and exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e with high sensitivity and minimal cytotoxicity in living cells. Distinct from conventional boronate-based probes, \u003cb\u003eLysoH\u003c/b\u003e\u003csub\u003e\u003cb\u003e2\u003c/b\u003e\u003c/sub\u003e\u003cb\u003eO\u003c/b\u003e\u003csub\u003e\u003cb\u003e2\u003c/b\u003e\u003c/sub\u003e employs a novel, biocompatible chemical transformation to achieve sensitive and selective monitoring of H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e within lysosomes.\u003c/p\u003e","manuscriptTitle":"Selective Lysosomal H2O2 – ROS Imaging with a Naphthalimide Probe Forming Hydroxylamine without Overoxidation","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-12-17 20:06:31","doi":"10.21203/rs.3.rs-8253737/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":"630d75ce-47c6-4a4d-979f-cb6fb7498791","owner":[],"postedDate":"December 17th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-12-25T09:54:25+00:00","versionOfRecord":[],"versionCreatedAt":"2025-12-17 20:06:31","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8253737","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8253737","identity":"rs-8253737","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}