A BODIPY-Based Probe Enables Fluorogenicity via Thiol-Dependent Modulation of Fluorophore Aggregation

Given the popular usage of BODIPY fluorophores in biological research, their propensity to aggregate in aqueous solution and impact their spectroscopic properties arguably warrants more attention. The probe under study herein serves as a case in point. A para-maleimide-substituted meso-phenyl BODIPY (p-MB) had previously been characterized in organic media, where its inherently high fluorescence ruled out its fluorogenic potential. Here, we have found that in aqueous solution, p-MB behaves differently, exhibiting a much-reduced fluorescence as a result of aggregation-caused quenching (ACQ). Additionally, p-MB is capable of responding to complementarily reactive substrates, including thiols and TCEP, to generate a substantial turn-on signal. The fluorescence restoration is largest when it reacts with those containing adjacent ionizable groups. By being part of a polar conjugate, p-MB assumes a disaggregated form, circumventing ACQ and unleashing up to similar to 1000-fold fluorescence enhancement through apparent disaggregation-induced emission (DIE). While our results support DIE as the turn-on mechanism, we found that the reactivity of the probe is much lower when it is given time to form stable aggregates. Therefore, contrary to the conventional depiction that a DIE probe works by dispersing from preformed aggregates to react with the target, our results suggest that it functions via a target-mediated inhibition of probe aggregation. Altogether, our work highlights the aggregation issue often faced by BODIPY-based probes and demonstrates how that can be exploited for turn-on sensing application. Furthermore, it reconstructs a different pathway for the DIE mechanism.

Automated summary

Given the popular usage of BODIPY fluorophores in biological research, it is necessary to pay more attention to their tendency to aggregate in aqueous solution and its impact on their spectroscopic properties. This study focuses on a particular probe and reveals that it exhibits a much-reduced fluorescence in aqueous solution due to aggregation-caused quenching (ACQ). However, it is capable of responding to complementary reactive substrates and generates a substantial turn-on signal. The probe assumes a disaggregated form and unleashes a significant fluorescence enhancement through disaggregation-induced emission (DIE). The results suggest that the probe functions via a target-mediated inhibition of probe aggregation instead of dispersing from preformed aggregates to react with the target.