Designing Red-Shifted Molecular Emitters Based on the Annulated Locked GFP Chromophore Derivatives

Bioimaging techniques require development of a wide variety of fluorescent probes that absorb and emit red light. One way to shift absorption and emission of a chromophore to longer wavelengths is to modify its chemical structure by adding polycyclic aromatic hydrocarbon (PAH) fragments, thus increasing the conjugation length of a molecule while maintaining its rigidity. Here, we consider four novel classes of conformationally locked Green Fluorescent Protein (GFP) chromophore derivatives obtained by extending their aromatic systems in different directions. Using high-level ab initio quantum chemistry calculations, we show that the alteration of their electronic structure upon annulation may unexpectedly result in a drastic change of their fluorescent properties. A flip of optically bright and dark electronic states is most prominent in the symmetric fluorene-based derivative. The presence of a completely dark lowest-lying excited state is supported by the experimentally measured extremely low fluorescence quantum yield of the newly synthesized compound. Importantly, one of the asymmetric modes of annulation provides a very promising strategy for developing red-shifted molecular emitters with an absorption wavelength of similar to 600 nm, having no significant impact on the character of the bright S-S-1 transition.

Automated summary

The development of bioimaging techniques relies on creating fluorescent probes that can absorb and emit red light, achieved by modifying the chemical structure of chromophores with polycyclic aromatic hydrocarbon fragments. Extending the conjugation length of a molecule can unexpectedly alter its electronic structure and fluorescent properties, leading to potential advancements in red-shifted molecular emitters for imaging applications.