Two-photon ratiometric fluorescent probe based on NBD-amine functionalized semiconducting polymer nanoparticles for real-time imaging of hydrogen sulfide in living cells and zebrafish

The thiolysis of 7-nitro-1,2,3-benzoxadiazole amine (NBD-A) paves the way for specific sensing of H 2 S over biothiols and real-time imaging in living organisms. Rational fabrication of NBD-A-based probe with ratiometric mode and two-photon excitation is highly appealing to achieve high quality bioimaging. In this work, the NBD-A molecules are assembled with poly(9,9-dioctylfluorenyl-2,7-diyl) polymer nanoparticles , defined as NBD@PFO, to construct two-photon ratiometric probes for H 2 S detection through the fluorescence resonance energy transfer (FRET). For the construction of NBD@PFO nanohybrids , polymer nanoparticles are employed as the NBD-A molecular vehicle, energy donor and two-photon absorber, while NBD-A is served as the response unit and energy acceptor. Taking advantages of NBD-A and polymer nanoparticles, the obtained NBD@PFO probes exhibit high selectivity, fast response (<5 s), ratiometric detection and two-photon excitation. Our results indicate that NBD@PFO nanohybrids are successfully applied for monitoring of H 2 S concentration in living cells and zebrafish, exhibiting great potential of polymer nanoparticles to improve the imaging capability of an organic small molecular probe .