Rigid, α-Helical Polypeptide Nanoprobes with Thermally Activated Delayed Fluorescence for Time-Resolved, High-Contrast Bioimaging

Thermally activated delayed fluorescence (TADF)-based nanoprobes are promising candidates as bioimaging agents, yet the fine-tuning of their photophysical properties through the modulation of the surrounding matrices remains largely unexplored. Herein, we report the development of polypeptide-TADF nanoprobes, where the rigid, α-helical polypeptide scaffold plays a critical role in enhancing the emission intensity and lifetime of the TADF fluorophore for bioimaging. The α-helical scaffolds not only spatially separated TADF molecules to avoid self-quenching but also anchored the dyes with minimized rotation and vibration. The nanoprobes thus exhibited >600 nm microsecond emission even in the presence of oxygen, facilitating cellular and animal imaging with a high signal-to-background ratio (SBR) by minimizing the interferences from autofluorescence signals. We believe that this work highlights the impact of the supporting polymeric conformation on the TADF performance, offering insights for the future design of time-resolved imaging probes.