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However, the application of fluorescence lifetime imaging microscopy (FLIM) for this purpose is hampered by the lack of probes with long fluorescence lifetimes and high signal\u2010to\u2010noise ratios. Herein, we developed two naphthalimide\u2010based dyed fluorescent probes, NBC and NBA, which are viscosity\u2010sensitive and feature donor\u2010\u03c0\u2010acceptor architectures. They operate via the restriction of intramolecular rotation in high\u2010viscosity environments exhibiting 10.7\/9.6\u2010fold fluorescence enhancement with strong linear correlations. In studies with HepG2 cells, both probes demonstrated excellent biocompatibility, specific targeting of lipid droplets, and remarkable photostability during prolonged imaging. FLIM analysis revealed dose\u2010dependent increases in lipid droplet viscosity following oleic acid treatment. The fluorescence lifetimes increased from 1.3 to 7.0\u00a0ns for NBA and from 2.5 to 8.2\u00a0ns for NBC. Time\u2010course experiments using the NBC probe captured distinct lipid droplet maturation stages, showing a progressive viscosity increase from newly formed droplets with a lifetime of 2.2\u00a0ns to mature droplets at 7.9\u00a0ns. These findings establish NBC and NBA as powerful tools for quantitative lipid droplet viscosity mapping, providing new insights into NAFLD pathogenesis and potentially enabling early diagnostic approaches for lipid metabolism disorders.<\/jats:p>","DOI":"10.1002\/asia.202500953","type":"journal-article","created":{"date-parts":[[2025,12,18]],"date-time":"2025-12-18T04:35:42Z","timestamp":1766032542000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Naphthalimide\u2010Based Dyed Long Fluorescent Lifetime Probe for Lipid Droplet Viscosity Mapping in Non\u2010Alcoholic Fatty Liver Disease"],"prefix":"10.1002","volume":"21","author":[{"given":"Yifan","family":"Liu","sequence":"first","affiliation":[{"name":"School of Chemistry and Chemical Engineering Beijing Institute of Technology  Beijing P. R. China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Feiran","family":"Liu","sequence":"additional","affiliation":[{"name":"School of Chemistry and Chemical Engineering Beijing Institute of Technology  Beijing P. R. China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tong","family":"Wu","sequence":"additional","affiliation":[{"name":"School of Chemistry and Chemical Engineering Beijing Institute of Technology  Beijing P. R. China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fei","family":"Yuan","sequence":"additional","affiliation":[{"name":"School of Chemistry and Chemical Engineering Beijing Institute of Technology  Beijing P. R. China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhiyang","family":"An","sequence":"additional","affiliation":[{"name":"School of Chemistry and Chemical Engineering Beijing Institute of Technology  Beijing P. R. 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