{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T05:05:14Z","timestamp":1789535114881,"version":"build-2803163510"},"posted":{"date-parts":[[2026,7,17]]},"group-title":"Cell Biology","reference-count":32,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,7,17]],"date-time":"2026-07-17T00:00:00Z","timestamp":1784246400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"name":"NIH","award":["R35GM153237"],"award-info":[{"award-number":["R35GM153237"]}]},{"name":"NIH","award":["R35GM149349"],"award-info":[{"award-number":["R35GM149349"]}]},{"name":"NIH","award":["F32GM139263"],"award-info":[{"award-number":["F32GM139263"]}]},{"name":"Agilent Biodesign Program"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,11]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  We report a generalizable method for single-molecule localization super-resolution microscopy in living cells. Live-cell PAINT (points accumulation for imaging in nanoscale topography) and single-molecule diffusivity mapping (SM\n                  <jats:italic>d<\/jats:italic>\n                  M) of intracellular protein targets are achieved by pairing exceptionally fluorogenic bis-trifluoromethyl rhodamine (BF) dyes with reversible HaloTag ligands and widely available HaloTag fusion proteins. This far-red small-molecule update to protein-based PAINT is readily incorporated into existing super-resolution microscopy workflows: wash-free staining and imaging are achieved for cell lines and primary neuron cultures for diverse protein targets. Pairing with photoconvertible fluorescent proteins further enables simultaneous two-color live-cell super-resolution microscopy and SM\n                  <jats:italic>d<\/jats:italic>\n                  M.\n                <\/jats:p>","DOI":"10.64898\/2026.07.16.738981","type":"posted-content","created":{"date-parts":[[2026,7,17]],"date-time":"2026-07-17T15:15:19Z","timestamp":1784301319000},"source":"Crossref","is-referenced-by-count":0,"title":["Live-cell PAINT microscopy of protein targets\n                  <i>via<\/i>\n                  fluorogenic exchangeable HaloTag ligands"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5997-0445","authenticated-orcid":false,"given":"Tong","family":"Zhan","sequence":"first","affiliation":[{"name":"Departments of Chemistry, University of California, Berkeley 94720-1460"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3991-1528","authenticated-orcid":false,"given":"Nels C.","family":"Gerstner","sequence":"additional","affiliation":[{"name":"Departments of Chemistry, University of California, Berkeley 94720-1460"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8797-0867","authenticated-orcid":false,"given":"Julia G.","family":"Martin","sequence":"additional","affiliation":[{"name":"Departments of Chemistry, University of California, Berkeley 94720-1460"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8812-829X","authenticated-orcid":false,"given":"Jack T.","family":"McCann","sequence":"additional","affiliation":[{"name":"Departments of Chemistry, University of California, Berkeley 94720-1460"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-4411-8844","authenticated-orcid":false,"given":"Marissa C.","family":"Lynch","sequence":"additional","affiliation":[{"name":"Departments of Chemistry, University of California, Berkeley 94720-1460"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-8502-2203","authenticated-orcid":false,"given":"Hsin-Jung","family":"Wu","sequence":"additional","affiliation":[{"name":"Departments of Chemistry, University of California, Berkeley 94720-1460"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2788-194X","authenticated-orcid":false,"given":"Ke","family":"Xu","sequence":"additional","affiliation":[{"name":"Departments of Chemistry, University of California, Berkeley 94720-1460"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6556-7679","authenticated-orcid":false,"given":"Evan W.","family":"Miller","sequence":"additional","affiliation":[{"name":"Departments of Chemistry, University of California, Berkeley 94720-1460"},{"name":"Departments of Molecular & Cell Biology, University of California, Berkeley 94720-1460"},{"name":"Departments of Helen Wills Neuroscience Institute, University of California, Berkeley 94720-1460"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026091521401090000_2026.07.16.738981v2.1","doi-asserted-by":"crossref","unstructured":"Xu, K. ; Shim, S.-H. ; Zhuang, X. , Super-resolution imaging through stochastic switching and localization of single molecules: an overview. In Far-Field Optical Nanoscopy, Tinnefeld, P. ; Eggeling, C. ; Hell, S. W. , Eds. Springer: Berlin, 2015; pp 27\u201364.","DOI":"10.1007\/4243_2013_61"},{"key":"2026091521401090000_2026.07.16.738981v2.2","doi-asserted-by":"publisher","DOI":"10.1038\/nrm.2017.71"},{"key":"2026091521401090000_2026.07.16.738981v2.3","doi-asserted-by":"publisher","DOI":"10.1126\/science.aau1044"},{"issue":"1","key":"2026091521401090000_2026.07.16.738981v2.4","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1038\/s43586-021-00038-x","article-title":"Single-molecule localization microscopy","volume":"1","year":"2021","journal-title":"Nat. Rev. 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