Molecular design strategy to alleviate environmental interference on two-photon fluorescence probes

Abstract

Electron donor-acceptor (D-A)-type molecules are the most widely employed two-photon scaffolds. Unfortunately, current D-A-type fluorophores suffer from serious solvent-dependent emission wavelength and brightness, thwarting their use for high-fidelity imaging in complicated biological systems. Herein, with single-atom replacement and acceptor fusing in acedan (6-acetyl-2-(dimethylamino)naphthalene), we devise a new class of D-A type fluorophore (TPQL dyes). Our design results in TPQL fluorophores with a slight emission redshift (Δλ TPQL1 < 32 nm versus Δλ Acedan-NH2 = 91 nm) and high brightness (εΦ TPQL1 = 7,600 to 6,923 versus εΦ Acedan-NH2 = 7,372 to 1,314) in different solvent systems. Such features not only provide bright and unbiased fluorescent images of TPQL fluorophores in the cell but also bring deeper tissue penetration and greater signal-to-background ratios in tissue. As a proof-of-concept demonstration, using probe TPQL-N 3 and TPQL-APN, we monitor the dynamic changes of H 2 S and APN during zebrafish liver development with high fidelity to the best of our knowledge. A new class of D-A-type fluorophore can alleviate environmental interference Decreasing dipole moment and suppressing TICT to inhibit solvent polarization Bright and unbiased fluorescent images in cells and tissues High-fidelity imaging of H 2 S and APN during zebrafish liver development Zhang et al. report a new class of D-A-type fluorophore (TPQL dyes), which not only displays solvent-insensitive fluorescence but also has high brightness in water. TPQL dyes are shown to be useful fluorophore scaffolds for the development of high-fidelity two-photon molecular probes in the biological research.