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However, understanding their cellular uptake and distribution and quantifying their presence at the single cell level remains challenging. Advanced imaging techniques, including transmission electron microscopy, synchrotron radiation X-ray fluorescence, and energetic ion beam-based nuclear microscopy (scanning transmission ion microscopy, particle-induced X-ray emission, elastic backscattering spectrometry), allow detailed high-resolution visualization of structure and morphology, high sensitivity for elemental detection with quantification within single cells, and the construction of 3D models of metal distribution, positioning them as powerful tools for assessing the cellular uptake and compartmentalization of complexes. Three Cu(II) complexes [Cu(phen)2(H2O)](NO3)2 (1), [Cu(Me2phen)2(NO3)]NO3 (2) and [Cu(amphen)2(H2O)](NO3)2 (3), (phen = 1,10-phenanthroline, Me2phen = 4,7-dimethyl-1,10-phen, amphen = 5-amino-phen) were investigated for Cu uptake and distribution in PC3 prostate cancer cells. All complexes show significant Cu uptake regardless of media concentration. Cu concentrations in the cytoplasm and nucleus are similar between treatments. Complexes 1 and 3 concentrate Cu in the nuclear region and show a vesicle-like pattern around the nucleus, while 2 shows a dispersed cytoplasmic pattern with large vesicles. The 3D models confirm that Cu is not retained at the plasma membrane, with complex 1 targeting the nucleus and 2 remaining in the cytoplasm. These results highlight the importance of quantifying metal distribution and correlating it with structural changes to understand the relevance of the ligand in the mechanisms of cellular uptake and targeting, crucial for the development of effective metal-based cancer therapies.<\/jats:p>","DOI":"10.1093\/mtomcs\/mfae051","type":"journal-article","created":{"date-parts":[[2024,11,8]],"date-time":"2024-11-08T03:37:09Z","timestamp":1731037029000},"source":"Crossref","is-referenced-by-count":5,"title":["Cellular targets of cytotoxic copper phenanthroline complexes: a multimodal imaging quantitative approach in single PC3 cells"],"prefix":"10.1093","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1836-2603","authenticated-orcid":false,"given":"Teresa","family":"Pinheiro","sequence":"first","affiliation":[{"name":"Instituto de Bioengenharia e Bioci\u00eancias, Departamento de Engenharia e Ci\u00eancias Nucleares, Instituto Superior T\u00e9cnico, Universidade de Lisboa , Av. Rovisco Pais 1, 1049-001 Lisboa ,","place":["Portugal"]}]},{"given":"Lu\u00eds C","family":"Alves","sequence":"additional","affiliation":[{"name":"Centro de Ci\u00eancias e Tecnologias Nucleares, Departamento de Engenharia e Ci\u00eancias Nucleares, Instituto Superior T\u00e9cnico, Universidade de Lisboa , Estrada Nacional 10, 2695-066 Bobadela LRS ,","place":["Portugal"]}]},{"given":"Ant\u00f3nio P","family":"Matos","sequence":"additional","affiliation":[{"name":"Egas Moniz Center for Interdisciplinary Research, Egas Moniz School of Health and Science, Campus Universit\u00e1rio, Quinta da Granja, Monte de Caparica , 2829-511 Caparica ,","place":["Portugal"]}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7096-4284","authenticated-orcid":false,"given":"Isabel","family":"Correia","sequence":"additional","affiliation":[{"name":"Centro de Qu\u00edmica Estrutural, Institute of Molecular Sciences, and Departamento de Engenharia Qu\u00edmica, Instituto Superior T\u00e9cnico, Universidade de Lisboa , Av. Rovisco Pais, 1049-001 Lisboa ,","place":["Portugal"]}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3978-9964","authenticated-orcid":false,"given":"Jo\u00e3o","family":"Costa Pessoa","sequence":"additional","affiliation":[{"name":"Centro de Qu\u00edmica Estrutural, Institute of Molecular Sciences, and Departamento de Engenharia Qu\u00edmica, Instituto Superior T\u00e9cnico, Universidade de Lisboa , Av. 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