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A panel of novel tridentate thiosemicarbazone ligands and their corresponding lipophilic and monocationic complexes with 1:2 metal\u2010to\u2010ligand stoichiometry was synthesized. These compounds are structurally related to clinically relevant thiosemicarbazone\u2010based systems with imaging and diagnostic applications. Comprehensive analytical characterization confirmed these probes\u2019 identity, while their complex solution behavior was evaluated under biologically relevant conditions. Cellular uptake and subcellular distribution were investigated in HeLa and PC\u20103 cells using correlated confocal microscopy and MP\u2010FLIM, revealing preferential mitochondrial localization for selected gallium complexes. In vivo imaging in zebrafish further unraveled the biodistribution of these fluorescent metal complexes, including selective labelling of neuromast structures. The combined in vitro and in vivo imaging data provide direct insight into the speciation, stability, and spatial distribution of thiosemicarbazone metal complexes in complex biological systems. This work establishes new fluorescent thiosemicarbazones as versatile platforms for studying metal complex behavior and biodistribution by advanced optical imaging techniques, opening new opportunities for advanced development of new theranostic applications.<\/jats:p>","DOI":"10.1002\/advs.202519815","type":"journal-article","created":{"date-parts":[[2026,4,16]],"date-time":"2026-04-16T19:21:41Z","timestamp":1776367301000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Mitochondria and Neuromast Tagging With Fluorescent Gallium\u2010Triapine Analogues: In Cellulo MP FLIM and Zebrafish Live Imaging"],"prefix":"10.1002","author":[{"given":"Megan J.","family":"Green","sequence":"first","affiliation":[{"name":"Department of Chemistry University of Bath Claverton Down  Bath UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael W.","family":"Jones","sequence":"additional","affiliation":[{"name":"Chemistry Research Laboratory University of Oxford  Oxford UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Merissa","family":"Saleem","sequence":"additional","affiliation":[{"name":"Department of Life Sciences University of Bath Claverton Down  Bath UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haobo","family":"Ge","sequence":"additional","affiliation":[{"name":"Department of Chemistry University of Bath Claverton Down  Bath UK"},{"name":"Department of Life Sciences University of Bath Claverton Down  Bath UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melita","family":"Tvardauskaite","sequence":"additional","affiliation":[{"name":"Department of Life Sciences University of Bath Claverton Down  Bath UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julia","family":"Dudzic","sequence":"additional","affiliation":[{"name":"Department of Life Sciences University of Bath Claverton Down  Bath UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fernando","family":"Cortezon\u2010Tamarit","sequence":"additional","affiliation":[{"name":"Department of Chemistry University of Bath Claverton Down  Bath UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rory L.","family":"Arrowsmith","sequence":"additional","affiliation":[{"name":"Department of Chemistry University of Bath Claverton Down  Bath UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charareh","family":"Pourzand","sequence":"additional","affiliation":[{"name":"Department of Life Sciences University of Bath Claverton Down  Bath UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriele","family":"Kociock\u2010Kohn","sequence":"additional","affiliation":[{"name":"Department of Chemistry University of Bath Claverton Down  Bath UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicholas H.","family":"Rees","sequence":"additional","affiliation":[{"name":"Chemistry Research Laboratory University of Oxford  Oxford UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonathan R.","family":"Dilworth","sequence":"additional","affiliation":[{"name":"Chemistry Research Laboratory University of Oxford  Oxford UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen","family":"Faulkner","sequence":"additional","affiliation":[{"name":"Chemistry Research Laboratory University of Oxford  Oxford UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stanley W.","family":"Botchway","sequence":"additional","affiliation":[{"name":"Central Laser Facility Rutherford Appleton Laboratory Research Complex at Harwell STFC  Didcot UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"Gurevich","sequence":"additional","affiliation":[{"name":"Department of Life Sciences University of Bath Claverton Down  Bath UK"},{"name":"Centre For Bioengineering &amp; Biomedical Technologies (CBio) University of Bath  Bath UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6385-4650","authenticated-orcid":false,"given":"Sofia I.","family":"Pascu","sequence":"additional","affiliation":[{"name":"Department of Chemistry University of Bath Claverton Down  Bath UK"},{"name":"Centre For Bioengineering &amp; Biomedical Technologies (CBio) University of Bath  Bath UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,4,16]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.3389\/fchem.2024.1424022"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms241512010"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00280\u2010016\u20103200\u2010x"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ygyno.2025.03.007"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/bjc.2016.54"},{"key":"e_1_2_8_7_1","doi-asserted-by":"crossref","first-page":"704","DOI":"10.1039\/B919119B","article-title":"Fluorescence Properties and Cellular Distribution of the Investigational Anticancer Drug Triapine (3\u2010Aminopyridine\u20102\u2010Carboxaldehyde Thiosemicarbazone) and Its Zinc(II) Complex","volume":"39","author":"Kowol C. 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