Highly Sensitive Fluorescence Probes for Nitric Oxide Based on Boron Dipyrromethene ChromophoreRational Design of Potentially Useful Bioimaging Fluorescence Probe

TLDR

Boron dipyrromethene (BODIPY) is a highly fluorescent chromophore in aqueous solution that has been underused for biological imaging compared to fluorescein. The authors created DAMBO‑PH, a BODIPY‑based probe that sensitively detects nitric oxide (NO) through a fluorescence turn‑on response. Fluorescence switching is achieved by an intramolecular photoinduced electron transfer (PeT) mechanism, engineered by tuning BODIPY electronic properties and adding hydrophilic groups to suppress pH dependence and aggregation quenching. Upon reacting with NO, DAMBO‑PH’s quantum yield rises from 0.002 to 0.74, illustrating the method’s promise for designing other bioimaging fluorescence probes.

Abstract

Boron dipyrromethene (BODIPY) is known to have a high quantum yield (φ) of fluorescence in aqueous solution but has not been utilized much for biological applications, compared to fluorescein. We developed 8-(3,4-diaminophenyl)-2,6-bis(2-carboxyethyl)-4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene (DAMBO-PH), based on the BODIPY chromophore, as a highly sensitive fluorescence probe for nitric oxide (NO). DAMBO-PH had a low φ value of 0.002, whereas its triazole derivative (DAMBO-PH-T), the product of the reaction of DAMBO-PH with NO, fluoresced strongly (φ = 0.74). The change of the fluorescence intensity was found to be controlled by an intramolecular photoinduced electron transfer (PeT) mechanism. The strategy for development of DAMBO-PH was as follows: (1) in order to design a highly sensitive probe of NO, the reactivity of o-phenylenediamine derivatives as NO-reactive moieties was examined using 4,5-diaminofluorescein (DAF-2, a widely used NO fluorescence probe), (2) in order to avoid pH-dependency of the fluorescence intensity, the PeT process was controlled by modulating the spectroscopic and electrochemical properties of BODIPY chromophores according to the Rehm−Weller equation based on measurement of excitation energies of chromophores, ground-state reduction potentials of PeT acceptors (BODIPYs), and calculation of the HOMO energy level of the PeT donor (o-phenylenediamine moiety) at the B3LYP/6-31G* level, (3) in order to avoid quenching of fluorescence by stacking of the probes and to obtain probes suitable for biological applications, hydrophilic functional groups were introduced. This strategy should be applicable for the rational design of other novel and potentially useful bioimaging fluorescence probes.