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The methodology yields Pdots with a high cationic surface charge (+56.5\u00a0mV \u00b1 9.5) and average hydrodynamic radius of 12\u00a0nm. Optical characterization reveals that these Pdots emit near\u2010infrared (NIR) light at a maximum wavelength of 860\u00a0nm owing to their conjugated polymer backbone consisting of D\u2013A\u2013D monomers. Both colloidal and optical properties of these Pdots make them promising fluorescence emissive probes for bioimaging applications. The significant advantage of positively charged Pdots is demonstrated in diffusion\u2010limited mediums such as tissues, utilizing human epithelial breast adenocarcinoma, ATCC HTB\u201022 (MCF\u20107), human bone marrow neuroblastoma, ATCC CRL\u20102266 (SH\u2010SY5Y), and rat adrenal gland pheochromocytoma, CRL\u20101721 (PC\u201012)\u00a0tumor spheroid models. Fluorescence microscopy analysis of tumor spheroids from MCF\u20107, SH\u2010SY5Y, and PC\u201012 cell lines reveals the intensity profile of Pdots, confirming extensive penetration into the central regions of the models. Moreover, a comparison with mitochondria staining dye reveals an overlap between the regions stained by Pdots and the dye in all three tumor spheroid models. These results suggest that single\u2010chain D\u2013A\u2013D type Pdots, cationized via CTAB, exhibit long\u2010range mean free path of penetration (\u22481\u00a0\u00b5m) in dense mediums and tumors.<\/jats:p>","DOI":"10.1002\/advs.202403398","type":"journal-article","created":{"date-parts":[[2024,7,18]],"date-time":"2024-07-18T08:03:08Z","timestamp":1721289788000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Near\u2010Infrared Emissive Super Penetrating Conjugated Polymer Dots for Intratumoral Imaging in 3D Tumor Spheroid Models"],"prefix":"10.1002","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3147-8627","authenticated-orcid":false,"given":"Soner","family":"Karabacak","sequence":"first","affiliation":[{"name":"Department of Chemistry Izmir Institute of Technology  Urla Izmir 35430 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