RESUMEN
Fluorescence imaging of living cells is key to better understanding cellular morphology and biological processes. Water-dispersible nanoparticles exhibiting thermally activated delayed fluorescence (TADF) have recently emerged as useful probes for time-resolved fluorescence imaging (TRFI), circumventing interference from biological autofluorescence. Many existing approaches, however, require TADF dyes with specific structural features, precluding many high-performance TADF materials from being used in this application. Here, we describe the synthesis of two TADF emitters based on the rigid and strongly electron-withdrawing dibenzo[a,c]dipyrido[3,2-h:2'-3'-j]phenazine-12-yl (BPPZ) motif, and demonstrate two parallel approaches for the encapsulation of these fluorophores to yield water-dispersible nanoparticles suitable for TRFI. First, fluorescent polymer dots (Pdots) were formed by dye encapsulation within cell-penetrating amphiphilic copolymers. Glassy organic nanoparticles (g-Odots) were also prepared, giving nanoparticles with higher photoluminescence quantum yields and improved colour purity. Both approaches yielded nanoparticles suitable for imaging, with reasonable uptake and cytotoxicity on the timescale of standard imaging experiments using human cervical (HeLa) and liver (HepG2) cancer cell lines. This work demonstrates two flexible strategies for preparing water-dispersible TADF nanoparticles for TRFI, both of which should be readily adaptable to nearly any existing hydrophobic TADF dye.