Glutamate indicators with increased sensitivity and tailored deactivation rates

Glutamate indicators with increased sensitivity and tailored deactivation rates | 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 Glutamate indicators with increased sensitivity and tailored deactivation rates Kaspar Podgorski, Abhi Aggarwal, Adrian Negrean, Yang Chen, Rishyashring Iyer, and 29 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6257403/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 23 Dec, 2025 Read the published version in Nature Methods → Version 1 posted You are reading this latest preprint version Abstract Identifying the input-output operations of neurons requires measurements of synaptic transmission simultaneously at many of a neuron’s thousands of inputs in the intact brain. To facilitate this goal, we engineered and screened 3365 variants of the fluorescent protein glutamate indicator iGluSnFR3 in neuron culture, and selected variants in the mouse visual cortex. Two variants have high sensitivity, fast activation (< 2 ms) and deactivation times tailored for recording large populations of synapses (iGluSnFR4s, 153 ms) or rapid dynamics (iGluSnFR4f, 26 ms). By imaging action-potential evoked signals on axons and visually-evoked signals on dendritic spines, we show that iGluSnFR4s/4f primarily detect local synaptic glutamate with single-vesicle sensitivity. The indicators detect a wide range of naturalistic synaptic transmission, including in the vibrissal cortex layer 4 and in hippocampal CA1 dendrites. iGluSnFR4 increases the sensitivity and scale (4s) or speed (4f) of tracking information flow in neural networks in vivo. Biological sciences/Neuroscience/Synaptic transmission Biological sciences/Biological techniques/Sensors and probes/Fluorescent proteins Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SupplementaryVideo1minislandscapebioRxiv.mp4 Supplementary Video 1: Example optical mini recording for iGluSnFR3, iGluSnFR4f, and iGluSnFR4s. Cyan denotes structure, and is the raw recording low pass filtered at 4 Hz, then gamma corrected (0.5). The residual (i.e. high-pass filtered recording) is shown in red (no gamma correction). Look up table for both channels are matched across the three videos. Video is played at 0.5x of the original recordings. SupplementaryVideo2vS1L4axons.mp4 Supplementary Video 2: Average 2P-AO recordings of evoked thalamocortical bouton activity (mean of 60 one-second intervals) in vS1 Layer 4 at 5 Hz stimulus frequency. Cite Share Download PDF Status: Published Journal Publication published 23 Dec, 2025 Read the published version in Nature Methods → 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-6257403","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":438611534,"identity":"38fe45bf-cbd4-48e0-8fb1-acde407126fe","order_by":0,"name":"Kaspar Podgorski","email":"data:image/png;base64,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","orcid":"https://orcid.org/0000-0002-0374-2005","institution":"Allen Institute for Neural Dynamics","correspondingAuthor":true,"prefix":"","firstName":"Kaspar","middleName":"","lastName":"Podgorski","suffix":""},{"id":438611535,"identity":"1cb52347-14aa-4b10-bea9-b47ba97c369d","order_by":1,"name":"Abhi Aggarwal","email":"","orcid":"https://orcid.org/0000-0002-6336-566X","institution":"Allen Institute for Neural Dynamics","correspondingAuthor":false,"prefix":"","firstName":"Abhi","middleName":"","lastName":"Aggarwal","suffix":""},{"id":438611536,"identity":"ac7be528-5d1d-48d8-b1b8-d69c2fe0e72f","order_by":2,"name":"Adrian Negrean","email":"","orcid":"https://orcid.org/0000-0002-9256-7412","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Adrian","middleName":"","lastName":"Negrean","suffix":""},{"id":438611537,"identity":"721ea82a-88af-4132-9d57-86867c88f24d","order_by":3,"name":"Yang Chen","email":"","orcid":"","institution":"TUM","correspondingAuthor":false,"prefix":"","firstName":"Yang","middleName":"","lastName":"Chen","suffix":""},{"id":438611538,"identity":"d0b3012a-cc13-4910-9cb2-3a051ebcc799","order_by":4,"name":"Rishyashring Iyer","email":"","orcid":"https://orcid.org/0000-0001-9126-9491","institution":"University of California San Diego","correspondingAuthor":false,"prefix":"","firstName":"Rishyashring","middleName":"","lastName":"Iyer","suffix":""},{"id":438611539,"identity":"c228435f-4563-47a9-868f-456eaac9818e","order_by":5,"name":"Daniel Reep","email":"","orcid":"https://orcid.org/0009-0009-7638-9538","institution":"Janelia Research Campus","correspondingAuthor":false,"prefix":"","firstName":"Daniel","middleName":"","lastName":"Reep","suffix":""},{"id":438611540,"identity":"5f184cb1-ddf2-488b-a85f-5df32450b329","order_by":6,"name":"Anyi Liu","email":"","orcid":"","institution":"University College London, Gower St, London WC1E 6BT, United Kingdom","correspondingAuthor":false,"prefix":"","firstName":"Anyi","middleName":"","lastName":"Liu","suffix":""},{"id":438611541,"identity":"dee1e0d7-34bc-4ece-8ecc-c22eebc03ed8","order_by":7,"name":"Anirudh Palutla","email":"","orcid":"","institution":"Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA 20147","correspondingAuthor":false,"prefix":"","firstName":"Anirudh","middleName":"","lastName":"Palutla","suffix":""},{"id":438611542,"identity":"8f969d80-8254-4399-8431-cd6273e5d6d0","order_by":8,"name":"Michael Xie","email":"","orcid":"https://orcid.org/0000-0002-4026-9181","institution":"Johns Hopkins University, Baltimore, MD, USA 21218","correspondingAuthor":false,"prefix":"","firstName":"Michael","middleName":"","lastName":"Xie","suffix":""},{"id":438611543,"identity":"b10965f0-80a9-41bc-905c-254e674573fd","order_by":9,"name":"Bryan MacLennan","email":"","orcid":"","institution":"Allen Institute for Neural Dynamics, Seattle, Washington, USA 98109","correspondingAuthor":false,"prefix":"","firstName":"Bryan","middleName":"","lastName":"MacLennan","suffix":""},{"id":438611544,"identity":"46eed355-1b45-4a71-9d2f-0cb7b4dfc4aa","order_by":10,"name":"Kenta Hagihara","email":"","orcid":"","institution":"Allen Institute for Neural Dynamics, Seattle, Washington, USA 98109","correspondingAuthor":false,"prefix":"","firstName":"Kenta","middleName":"","lastName":"Hagihara","suffix":""},{"id":438611545,"identity":"0a1022e9-1d94-4757-b3cc-655b28b22c28","order_by":11,"name":"Lucas Kinsey","email":"","orcid":"","institution":"Allen Institute for Neural Dynamics, Seattle, Washington, USA 98109","correspondingAuthor":false,"prefix":"","firstName":"Lucas","middleName":"","lastName":"Kinsey","suffix":""},{"id":438611546,"identity":"f7e4eb64-827d-4bdb-9bf0-7b95ba494181","order_by":12,"name":"Julianna Sun","email":"","orcid":"","institution":"Neuronal Cell Biology Division, Department of Developmental Neurobiology, St. Jude Children’s Research Hospital, 262 Danny Thomas Place, Memphis, TN, USA 38104","correspondingAuthor":false,"prefix":"","firstName":"Julianna","middleName":"","lastName":"Sun","suffix":""},{"id":438611547,"identity":"1486113a-c801-41db-8b51-0bd0cab296d1","order_by":13,"name":"Pantong Yao","email":"","orcid":"","institution":"Department of Neurosciences, University of California, San Diego, La Jolla, California, USA 92093","correspondingAuthor":false,"prefix":"","firstName":"Pantong","middleName":"","lastName":"Yao","suffix":""},{"id":438611548,"identity":"e4e34c30-795e-42e7-b918-f806c970dce2","order_by":14,"name":"Jihong Zheng","email":"","orcid":"https://orcid.org/0000-0002-1247-6087","institution":"Janelia Research Campus, Howard Hughes Medical Institute","correspondingAuthor":false,"prefix":"","firstName":"Jihong","middleName":"","lastName":"Zheng","suffix":""},{"id":438611549,"identity":"8802dd81-c9a7-4081-b5a7-365cadf66cb6","order_by":15,"name":"Arthur Tsang","email":"","orcid":"","institution":"Howard Hughes Medical Institute","correspondingAuthor":false,"prefix":"","firstName":"Arthur","middleName":"","lastName":"Tsang","suffix":""},{"id":438611550,"identity":"ead14909-fcf7-4273-9ab8-4b882c37fdb6","order_by":16,"name":"Getahun Tsegaye","email":"","orcid":"","institution":"Howard Hughes Medical Institute","correspondingAuthor":false,"prefix":"","firstName":"Getahun","middleName":"","lastName":"Tsegaye","suffix":""},{"id":438611551,"identity":"06310143-b106-448b-b109-eaf0988fac8c","order_by":17,"name":"Yonghai Zhang","email":"","orcid":"https://orcid.org/0009-0004-6121-427X","institution":"Institute of Neuroscience and Munich Cluster for Systems Neurology, Technical University of Munich, Munich, Germany.","correspondingAuthor":false,"prefix":"","firstName":"Yonghai","middleName":"","lastName":"Zhang","suffix":""},{"id":438611552,"identity":"06760149-289f-4a8d-910a-3b2c5a266c42","order_by":18,"name":"Ronak Patel","email":"","orcid":"","institution":"Howard Hughes Medical Institute","correspondingAuthor":false,"prefix":"","firstName":"Ronak","middleName":"","lastName":"Patel","suffix":""},{"id":438611553,"identity":"e9356bf5-3ea0-4475-9f49-e18c37b952fb","order_by":19,"name":"Benjamin Arthur","email":"","orcid":"https://orcid.org/0000-0003-3545-8807","institution":"Janelia Research Campus, Howard Hughes Medical Institute","correspondingAuthor":false,"prefix":"","firstName":"Benjamin","middleName":"","lastName":"Arthur","suffix":""},{"id":438611554,"identity":"5669b3e7-fffd-4cc4-be26-b5111211a8d8","order_by":20,"name":"Julien Hiblot","email":"","orcid":"","institution":"Max Planck Institute for Medical Research, Heidelberg, Germany, 69120","correspondingAuthor":false,"prefix":"","firstName":"Julien","middleName":"","lastName":"Hiblot","suffix":""},{"id":438611555,"identity":"c57e9aaf-2792-4463-b649-889d82bb6422","order_by":21,"name":"Philipp Leippe","email":"","orcid":"","institution":"CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Lazarettgasse 14, AKH BT 25.3, 1090 Wien, Austria","correspondingAuthor":false,"prefix":"","firstName":"Philipp","middleName":"","lastName":"Leippe","suffix":""},{"id":438611556,"identity":"9df6a574-9eb5-470e-87a5-b3d7557f1009","order_by":22,"name":"Miroslaw Tarnawski","email":"","orcid":"https://orcid.org/0000-0001-5297-5668","institution":"MPI for Medical Research","correspondingAuthor":false,"prefix":"","firstName":"Miroslaw","middleName":"","lastName":"Tarnawski","suffix":""},{"id":438611557,"identity":"6d7dff56-3f0b-4002-bb63-624c031150e1","order_by":23,"name":"Jonathan Marvin","email":"","orcid":"https://orcid.org/0000-0003-2294-4515","institution":"Howard Hughes Medical Institute, Janelia Research Campus","correspondingAuthor":false,"prefix":"","firstName":"Jonathan","middleName":"","lastName":"Marvin","suffix":""},{"id":438611558,"identity":"5085e411-a079-48df-9604-371f4c61cd28","order_by":24,"name":"Jason Vevea","email":"","orcid":"","institution":"Neuronal Cell Biology Division, Department of Developmental Neurobiology, St. Jude Children’s Research Hospital, 262 Danny Thomas Place, Memphis, TN, USA 38104","correspondingAuthor":false,"prefix":"","firstName":"Jason","middleName":"","lastName":"Vevea","suffix":""},{"id":438611559,"identity":"536ad35d-f038-43f2-b671-6eeb5a62cfb2","order_by":25,"name":"Srinivas Turaga","email":"","orcid":"https://orcid.org/0000-0003-3247-6487","institution":"HHMI Janelia Research Campus","correspondingAuthor":false,"prefix":"","firstName":"Srinivas","middleName":"","lastName":"Turaga","suffix":""},{"id":438611560,"identity":"ba363381-aa18-4524-8a52-107acd698c13","order_by":26,"name":"Alison Tebo","email":"","orcid":"https://orcid.org/0000-0003-0788-5617","institution":"HHMI Janelia","correspondingAuthor":false,"prefix":"","firstName":"Alison","middleName":"","lastName":"Tebo","suffix":""},{"id":438611561,"identity":"bb28a6a5-df36-4bef-8043-6e09d9092bfd","order_by":27,"name":"Matteo Carandini","email":"","orcid":"https://orcid.org/0000-0003-4880-7682","institution":"University College London","correspondingAuthor":false,"prefix":"","firstName":"Matteo","middleName":"","lastName":"Carandini","suffix":""},{"id":438611562,"identity":"a74879f1-a7eb-4f24-b988-231a8643b336","order_by":28,"name":"L. Rossi","email":"","orcid":"","institution":"Italian Institute of Technology,Naples, CAMPANIA, Italy, 80125","correspondingAuthor":false,"prefix":"","firstName":"L.","middleName":"","lastName":"Rossi","suffix":""},{"id":438611563,"identity":"56a6b528-624d-4ed7-baf4-87e001c0b05e","order_by":29,"name":"David Kleinfeld","email":"","orcid":"https://orcid.org/0000-0001-9797-4722","institution":"University of California San Diego","correspondingAuthor":false,"prefix":"","firstName":"David","middleName":"","lastName":"Kleinfeld","suffix":""},{"id":438611564,"identity":"a40e90ca-52fa-41e9-ae17-0b7a7a7d85fc","order_by":30,"name":"Arthur Konnerth","email":"","orcid":"https://orcid.org/0000-0002-9548-2676","institution":"Technical University of Munich","correspondingAuthor":false,"prefix":"","firstName":"Arthur","middleName":"","lastName":"Konnerth","suffix":""},{"id":438611565,"identity":"49c9c5b5-91f7-4aeb-9fc7-33637dda60a4","order_by":31,"name":"Karel Svoboda","email":"","orcid":"https://orcid.org/0000-0002-6670-7362","institution":"Allen Institute for Brain Science, Howard Hughes Medical Institute Janelia Research Campus","correspondingAuthor":false,"prefix":"","firstName":"Karel","middleName":"","lastName":"Svoboda","suffix":""},{"id":438611566,"identity":"9159ea69-022a-4c70-b286-015ee91cfcaf","order_by":32,"name":"Glenn Turner","email":"","orcid":"https://orcid.org/0000-0002-5341-2784","institution":"Janelia Research Campus, Howard Hughes Medical Institute","correspondingAuthor":false,"prefix":"","firstName":"Glenn","middleName":"","lastName":"Turner","suffix":""},{"id":438611567,"identity":"aafe1791-5309-482c-ad50-f533dd4dab85","order_by":33,"name":"Jeremy Hasseman","email":"","orcid":"https://orcid.org/0000-0002-0096-7321","institution":"Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA","correspondingAuthor":false,"prefix":"","firstName":"Jeremy","middleName":"","lastName":"Hasseman","suffix":""}],"badges":[],"createdAt":"2025-03-19 03:20:14","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6257403/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6257403/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41592-025-02965-z","type":"published","date":"2025-12-23T05:00:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":98928015,"identity":"551b9760-ad2e-4897-8dcf-103f84ae608e","added_by":"auto","created_at":"2025-12-24 08:11:32","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2881015,"visible":true,"origin":"","legend":"Article File","description":"","filename":"iGluSnFR4ManuscriptFinal.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6257403/v1_covered_eeefd34b-e326-447f-b206-cfc7dd5c2d5d.pdf"},{"id":80155414,"identity":"1dd52107-1b2b-4110-bd41-6f928c083ec1","added_by":"auto","created_at":"2025-04-08 14:21:04","extension":"mp4","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":189527508,"visible":true,"origin":"","legend":"\u003cp\u003eSupplementary Video 1: Example optical mini recording for iGluSnFR3, iGluSnFR4f, and iGluSnFR4s. Cyan denotes structure, and is the raw recording low pass filtered at 4 Hz, then gamma corrected (0.5). The residual (i.e. high-pass filtered recording) is shown in red (no gamma correction). Look up table for both channels are matched across the three videos. Video is played at 0.5x of the original recordings.\u003c/p\u003e","description":"","filename":"SupplementaryVideo1minislandscapebioRxiv.mp4","url":"https://assets-eu.researchsquare.com/files/rs-6257403/v1/d26abe40a1bb2c63bcfddbe1.mp4"},{"id":80155412,"identity":"d5686127-54ea-4314-8e07-66caa91a2a8c","added_by":"auto","created_at":"2025-04-08 14:21:00","extension":"mp4","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":8404116,"visible":true,"origin":"","legend":"\u003cp\u003eSupplementary Video 2: Average 2P-AO recordings of evoked thalamocortical bouton activity (mean of 60 one-second intervals) in vS1 Layer 4 at 5 Hz stimulus frequency.\u003c/p\u003e","description":"","filename":"SupplementaryVideo2vS1L4axons.mp4","url":"https://assets-eu.researchsquare.com/files/rs-6257403/v1/a76ee919a7d078e9951e93a8.mp4"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.","formattedTitle":"Glutamate indicators with increased sensitivity and tailored deactivation rates","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"nature-portfolio","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"","title":"Nature Portfolio","twitterHandle":"","acdcEnabled":false,"dfaEnabled":false,"editorialSystem":"ejp","reportingPortfolio":"","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-6257403/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6257403/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eIdentifying \u0026nbsp;the \u0026nbsp;input-output \u0026nbsp;operations \u0026nbsp;of \u0026nbsp;neurons \u0026nbsp;requires \u0026nbsp;measurements \u0026nbsp;of \u0026nbsp;synaptic transmission simultaneously at many of a neuron’s thousands of inputs in the intact brain. To facilitate this goal, we engineered and screened 3365 variants of the fluorescent protein glutamate indicator iGluSnFR3 in neuron culture, and selected variants in the mouse visual cortex. Two variants have high sensitivity, fast activation (\u0026lt; 2 ms) and deactivation times tailored for recording large populations of synapses (iGluSnFR4s, 153 ms) or rapid dynamics (iGluSnFR4f, 26 ms). By imaging action-potential evoked signals on axons and visually-evoked signals on dendritic spines, we \u0026nbsp;show \u0026nbsp;that \u0026nbsp;iGluSnFR4s/4f \u0026nbsp;primarily \u0026nbsp;detect \u0026nbsp;local \u0026nbsp;synaptic \u0026nbsp;glutamate \u0026nbsp;with \u0026nbsp;single-vesicle sensitivity. The indicators detect a wide range \u0026nbsp;of naturalistic synaptic transmission, including in the vibrissal cortex layer 4 and in hippocampal CA1 dendrites. iGluSnFR4 increases the sensitivity and scale (4s) or speed (4f) of tracking information flow in neural networks in vivo.\u0026nbsp;\u003c/p\u003e","manuscriptTitle":"Glutamate indicators with increased sensitivity and tailored deactivation rates","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-04-08 14:20:55","doi":"10.21203/rs.3.rs-6257403/v1","editorialEvents":[],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"nature-methods","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"nmeth","sideBox":"Learn more about [Nature Methods](http://www.nature.com/nmeth)","snPcode":"","submissionUrl":"","title":"Nature Methods","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature Research","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"980863d0-1e96-4b71-868a-b8b4d764d1b0","owner":[],"postedDate":"April 8th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":46708739,"name":"Biological sciences/Neuroscience/Synaptic transmission"},{"id":46708740,"name":"Biological sciences/Biological techniques/Sensors and probes/Fluorescent proteins"}],"tags":[],"updatedAt":"2025-12-24T08:11:24+00:00","versionOfRecord":{"articleIdentity":"rs-6257403","link":"https://doi.org/10.1038/s41592-025-02965-z","journal":{"identity":"nature-methods","isVorOnly":false,"title":"Nature Methods"},"publishedOn":"2025-12-23 05:00:00","publishedOnDateReadable":"December 23rd, 2025"},"versionCreatedAt":"2025-04-08 14:20:55","video":"","vorDoi":"10.1038/s41592-025-02965-z","vorDoiUrl":"https://doi.org/10.1038/s41592-025-02965-z","workflowStages":[]},"version":"v1","identity":"rs-6257403","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6257403","identity":"rs-6257403","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}