{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T17:20:40Z","timestamp":1787678440815,"version":"build-2784847793"},"reference-count":47,"publisher":"National Academy of Sciences","issue":"35","license":[{"start":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T00:00:00Z","timestamp":1787616000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002347","name":"Bundesministerium f\u00fcr Forschung, Technologie und Raumfahrt","doi-asserted-by":"publisher","award":["13N14122"],"award-info":[{"award-number":["13N14122"]}],"id":[{"id":"10.13039\/501100002347","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000781","name":"EC | European Research Council","doi-asserted-by":"publisher","award":["835102"],"award-info":[{"award-number":["835102"]}],"id":[{"id":"10.13039\/501100000781","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["DFG-CRC 1456 Project C06"],"award-info":[{"award-number":["DFG-CRC 1456 Project C06"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100018703","name":"EC | Horizon Europe | Innovative Europe | HORIZON EUROPE European Innovation Council","doi-asserted-by":"publisher","award":["848827"],"award-info":[{"award-number":["848827"]}],"id":[{"id":"10.13039\/100018703","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2026,9,1]]},"abstract":"<jats:p>The near-infrared (NIR) spectral region is attractive for live-cell imaging, due to low autofluorescence and reduced phototoxicity. Some phytochrome-derived fluorescent proteins absorb and emit fluorescence in the NIR, but have short fluorescence lifetimes and relatively low quantum yields, requiring higher laser powers thus limiting their usefulness for live-cell superresolution microscopy. Using the bacterial phytochrome miRFP703 as a template, we screened for variants with longer fluorescence lifetimes, because the quantum yield and fluorescence lifetime are linked. We identified the bright monomeric fluorescent protein elite-niRFP704, which has a longer lifetime (1.12 ns) and a correspondingly higher quantum yield (0.21) than its template, absorbing and emitting completely in the NIR spectral region. elite-niRFP704 was used to tag proteins in living cells and facilitated extended stimulated emission depletion (STED) microscopy on cell lines stably expressing a fusion protein. Finally, elite-niRFP704 and miRFP703 could be separated based on their significantly different lifetimes, enabling two-channel NIR STED microscopy of living mammalian cells.<\/jats:p>","DOI":"10.1073\/pnas.2533451123","type":"journal-article","created":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T16:36:09Z","timestamp":1787675769000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":0,"title":["Bright monomeric fluorescent protein elite-niRFP704 for two-channel near-infrared STED nanoscopy"],"prefix":"10.1073","volume":"123","author":[{"given":"Florian","family":"Habenstein","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03av75f26","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences","place":["G\u00f6ttingen, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nickels","family":"Jensen","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03av75f26","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences","place":["G\u00f6ttingen, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel","family":"Stumpf","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03av75f26","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences","place":["G\u00f6ttingen, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexey I.","family":"Chizhik","sequence":"additional","affiliation":[{"name":"III. Institute of Physics, Georg August University","place":["G\u00f6ttingen, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marcel","family":"Leutenegger","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03av75f26","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences","place":["G\u00f6ttingen, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1101-8516","authenticated-orcid":false,"given":"Jin","family":"Chang","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02e2c7k09","id-type":"ROR","asserted-by":"publisher"}],"name":"Optics Research Group, ImPhys Department, Faculty of Applied Sciences, Delft University of Technology","place":["Delft, The Netherlands"]},{"name":"Single Quantum B.V.","place":["Delft, The Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreas","family":"Fognini","sequence":"additional","affiliation":[{"name":"Single Quantum B.V.","place":["Delft, The Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessie","family":"Qin-Dregely","sequence":"additional","affiliation":[{"name":"Single Quantum B.V.","place":["Delft, The Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Iman Esmaeil","family":"Zadeh","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02e2c7k09","id-type":"ROR","asserted-by":"publisher"}],"name":"Optics Research Group, ImPhys Department, Faculty of Applied Sciences, Delft University of Technology","place":["Delft, The Netherlands"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5443-4344","authenticated-orcid":false,"given":"Laura L.","family":"Kirck","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03av75f26","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences","place":["G\u00f6ttingen, Germany"]},{"id":[{"id":"https:\/\/ror.org\/01s1h3j07","id-type":"ROR","asserted-by":"publisher"}],"name":"Fraunhofer Institute for Translational Medicine and Pharmacology ITMP, Translational Neuroinflammation and Automated Microscopy TNM","place":["G\u00f6ttingen, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5091-7157","authenticated-orcid":false,"given":"J\u00f6rg","family":"Enderlein","sequence":"additional","affiliation":[{"name":"III. Institute of Physics, Georg August University","place":["G\u00f6ttingen, Germany"]},{"id":[{"id":"https:\/\/ror.org\/01y9bpm73","id-type":"ROR","asserted-by":"publisher"}],"name":"Multiscale Bioimaging Cluster of Excellence, University of G\u00f6ttingen","place":["G\u00f6ttingen, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5959-6409","authenticated-orcid":false,"given":"Kaushik","family":"Inamdar","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03av75f26","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences","place":["G\u00f6ttingen, Germany"]},{"id":[{"id":"https:\/\/ror.org\/01y9bpm73","id-type":"ROR","asserted-by":"publisher"}],"name":"Clinic of Neurology, University of G\u00f6ttingen","place":["G\u00f6ttingen, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9638-5077","authenticated-orcid":false,"given":"Stefan W.","family":"Hell","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03av75f26","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences","place":["G\u00f6ttingen, Germany"]},{"id":[{"id":"https:\/\/ror.org\/01y9bpm73","id-type":"ROR","asserted-by":"publisher"}],"name":"Multiscale Bioimaging Cluster of Excellence, University of G\u00f6ttingen","place":["G\u00f6ttingen, Germany"]},{"id":[{"id":"https:\/\/ror.org\/000bxzc63","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Optical Nanoscopy, Max Planck Institute for Medical Research","place":["Heidelberg, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8028-3121","authenticated-orcid":false,"given":"Stefan","family":"Jakobs","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03av75f26","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences","place":["G\u00f6ttingen, Germany"]},{"id":[{"id":"https:\/\/ror.org\/01s1h3j07","id-type":"ROR","asserted-by":"publisher"}],"name":"Fraunhofer Institute for Translational Medicine and Pharmacology ITMP, Translational Neuroinflammation and Automated Microscopy TNM","place":["G\u00f6ttingen, Germany"]},{"id":[{"id":"https:\/\/ror.org\/01y9bpm73","id-type":"ROR","asserted-by":"publisher"}],"name":"Multiscale Bioimaging Cluster of Excellence, University of G\u00f6ttingen","place":["G\u00f6ttingen, Germany"]},{"id":[{"id":"https:\/\/ror.org\/01y9bpm73","id-type":"ROR","asserted-by":"publisher"}],"name":"Clinic of Neurology, University of G\u00f6ttingen","place":["G\u00f6ttingen, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2026,8,25]]},"reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibtech.2018.06.011"},{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/nm0103-123"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/86684"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrm.2017.71"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00038.2009"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-biochem-060614-034411"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-3040.1997.d01-108.x"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature04118"},{"key":"e_1_3_4_9_2","first-page":"273","volume-title":"Heme, Chlorophyll, and Bilins: Methods and Protocols","author":"Terry M. J.","year":"2002","unstructured":"M. J. Terry, \u201cBiosynthesis and analysis of bilins\u201d in Heme, Chlorophyll, and Bilins: Methods and Protocols, A. G. Smith, M. Witty, Eds. (Humana Press, Totowa, NJ, 2002), pp. 273-291."},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1168683"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.chemrev.6b00700"},{"key":"e_1_3_4_12_2","first-page":"32","article-title":"Near-infrared fluorescent proteins and their applications","volume":"84","author":"Karasev M. M.","year":"2019","unstructured":"M. M. Karasev, O. V. Stepanenko, K. A. Rumyantsev, K. K. Turoverov, V. V. Verkhusha, Near-infrared fluorescent proteins and their applications. Biochemistry 84, 32\u201350 (2019).","journal-title":"Biochemistry"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.3935"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-08050-8"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1021\/acssensors.2c00286"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1002\/pro.4709"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-13897-6"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-07246-2"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-3495(97)78307-3"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41592-023-01975-z"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms12405"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.202200267"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-38433-2"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41596-019-0250-7"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1415"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jpcb.2c01956"},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2025.08.035"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1039\/C7SC00855D"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1038\/s42003-020-01316-2"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1039\/C8OB02262C"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1111\/febs.12781"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1038\/srep37362"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.adm6869"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-11827-4"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-18640-z"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41592-026-03061-6"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-60761-652-8_7"},{"key":"e_1_3_4_38_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.82.2.488"},{"key":"e_1_3_4_39_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/28.16.e78"},{"key":"e_1_3_4_40_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.1952"},{"key":"e_1_3_4_41_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature10497"},{"key":"e_1_3_4_42_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0030-4018(96)00384-7"},{"key":"e_1_3_4_43_2","doi-asserted-by":"publisher","DOI":"10.1364\/AO.58.009803"},{"key":"e_1_3_4_44_2","doi-asserted-by":"publisher","DOI":"10.1038\/srep09592"},{"key":"e_1_3_4_45_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gku1082"},{"key":"e_1_3_4_46_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkt1361"},{"key":"e_1_3_4_47_2","unstructured":"K. Inamdar Bright monomeric fluorescent protein elite-niRFP704 for two-channel near- infrared STED nanoscopy. Zenodo. https:\/\/doi.org\/10.5281\/zenodo.21623096. Deposited 27 July 2026."}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.2533451123","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T16:37:05Z","timestamp":1787675825000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/10.1073\/pnas.2533451123"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,25]]},"references-count":47,"journal-issue":{"issue":"35","published-print":{"date-parts":[[2026,9,1]]}},"alternative-id":["10.1073\/pnas.2533451123"],"URL":"https:\/\/doi.org\/10.1073\/pnas.2533451123","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,25]]},"assertion":[{"value":"2025-11-19","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-06-29","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-08-25","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e2533451123"}}