{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T18:55:11Z","timestamp":1790794511004,"version":"4.1.0"},"reference-count":33,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"40","funder":[{"DOI":"10.13039\/100016394","name":"Ludwig Center at Harvard","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100016394","id-type":"DOI","asserted-by":"publisher"}]},{"name":"National Institutes of Health under Award Number","award":["K25EB032864"],"award-info":[{"award-number":["K25EB032864"]}]}],"content-domain":{"domain":["www.science.org"],"crossmark-restriction":true},"short-container-title":["Sci. Adv."],"published-print":{"date-parts":[[2026,10,2]]},"abstract":"<jats:p>Fully capturing the heterogeneity of biological processes remains a central challenge, as conventional confocal microscopy typically surveys large ensembles of molecules within a small volume. The development of super-resolution imaging enables the capture of images with exceptional detail. When super-resolution is combined with functional imaging, such as fluorescence lifetime and spectral imaging, it provides better separation of image constituents and sensing of environmental properties at the nanoscale. These multidimensional imaging approaches often acquire functional and fluorescence images separately and then merge the resulting datasets, a post hoc approach that risks dynamic range mismatches and the loss of subtle molecular signals. Here, we introduce a computational super-resolution framework based on the deblurring by pixel reassignment (DPR) algorithm, which processes multidimensional data as high-dimensional tensors, integrating spatial axes with fluorescence lifetime or spectral information into a single dataset. Using DPR, high-resolution details can be extracted from conventional fluorescence microscopy images, even from a single capture. We show that this approach enhances the spatial resolution of multidimensional datasets without compromising quantitative fidelity. As a proof of concept, we demonstrate super-resolution multimodal imaging (intensity, lifetime, and spectra), revealing fine-scale details while preserving signals from rare or low-intensity molecular events. This accessible yet powerful method paves the way for quantitative, single-molecule\u2013level insights into complex biological systems.<\/jats:p>","DOI":"10.1126\/sciadv.aec7970","type":"journal-article","created":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T17:58:26Z","timestamp":1790791106000},"update-policy":"https:\/\/doi.org\/10.34133\/aaas_crossmark","source":"Crossref","is-referenced-by-count":0,"title":["A computational super-resolution framework for multidimensional fluorescence imaging"],"prefix":"10.1126","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6156-9554","authenticated-orcid":true,"given":"Wonsang","family":"Hwang","sequence":"first","affiliation":[{"name":"Wellman Center for Photomedicine, Harvard Medical School, Massachusetts General Hospital, CNY149, 13th St, Charlestown, MA 02129, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-5208-5317","authenticated-orcid":true,"given":"Bingying","family":"Zhao","sequence":"additional","affiliation":[{"name":"Department of Electrical and Computer Engineering, Boston University, 8 St Mary\u2019s St, Boston, MA 02215, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kai","family":"Guo","sequence":"additional","affiliation":[{"name":"College of Graduate and Professional Studies, Trine University, 999 Republic Dr Suite 200, Allen Park, Detroit, MI 48101, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6676-0358","authenticated-orcid":true,"given":"Jenu V.","family":"Chacko","sequence":"additional","affiliation":[{"name":"Laboratory for Optical and Computational Instrumentation, University of Wisconsin-Madison, Madison, WI 53706, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9407-7006","authenticated-orcid":true,"given":"Sinyoung","family":"Jeong","sequence":"additional","affiliation":[{"name":"Intek Scientific, 1 Broadway, Cambridge, MA 02142, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4389-5516","authenticated-orcid":true,"given":"Ad\u00e1n","family":"Guerrero","sequence":"additional","affiliation":[{"name":"Laboratorio Nacional de Microscop\u00eda Avanzada, Instituto de Biotecnolog\u00eda, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Cuernavaca, Morelos, Mexico."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6343-9155","authenticated-orcid":true,"given":"Jerome","family":"Mertz","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, Boston University, 44 Cummington Mall, Boston, MA 02215, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2185-6505","authenticated-orcid":true,"given":"Conor L.","family":"Evans","sequence":"additional","affiliation":[{"name":"Wellman Center for Photomedicine, Harvard Medical School, Massachusetts General Hospital, CNY149, 13th St, Charlestown, MA 02129, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3611-7959","authenticated-orcid":true,"given":"Iv\u00e1n Coto","family":"Hern\u00e1ndez","sequence":"additional","affiliation":[{"name":"Center for Interdisciplinary Innovation in Imaging, Massachusetts General Hospital and Harvard Medical School, 149 13th St, Charlestown, MA 02129, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-22518-0"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-024-45567-7"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1117\/1.3494566"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.26508\/lsa.202302398"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1038\/s42003-024-06702-8"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1007\/s44352-025-00010-5"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1364\/OL.33.000113"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1021\/acsnano.0c07322"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41566-024-01481-4"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms12471"},{"key":"e_1_3_2_12_2","doi-asserted-by":"crossref","unstructured":"X. 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