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However, its application in living cells remains limited by the lack of suitable labeling strategies and the challenge of imaging dynamic cellular structures over extended periods. In this study, we introduce PAINTpro, a protein\u2010based PAINT technique that facilitates live\u2010cell SMLM imaging and visualizes dynamic structures at the nanoscale. PAINTpro utilizes an engineered reversible bilirubin (BR)\u2010binding protein, Fluorescent protein for Maximizing Imaging with Nano\u2010resolution (FluoroMIN), derived from the green fluorescent protein UnaG. FluoroMIN incorporates mutations based on in silico molecular dynamics simulations to accelerate BR exchange, resulting in reversible fluorescence, high single\u2010molecule signal yield, and prolonged switching lifetime. These properties enable both live\u2010cell labeling and super\u2010resolution PAINT imaging with a resolution of \u223c50\u00a0nm. Moreover, PAINTpro supports long\u2010term imaging of dynamic structures, providing time\u2010resolved super\u2010resolution images of microtubules with a resolution of up to 60\u00a0nm. Notably, PAINTpro is readily adaptable from existing protein\u2010PAINT protocols, offering a powerful tool for super\u2010resolution visualization of biological processes.<\/jats:p>","DOI":"10.1002\/smtd.70945","type":"journal-article","created":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T09:25:56Z","timestamp":1786440356000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["PAINTpro: Protein\u2010Based PAINT in Living Cells Using Engineered Reversible Bilirubin\u2010Binding Fluorescent Protein"],"prefix":"10.1002","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8754-0805","authenticated-orcid":false,"given":"Min","family":"Zhu","sequence":"first","affiliation":[{"name":"Department of Biomedical Engineering, State Key Laboratory of Extreme Photonics and Instrumentation, Key Laboratory of Biomedical Engineering of Ministry of Education, Zhejiang Key Laboratory of Intelligent Sensing Technology and Advanced 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