Co-aggregation as A Simple Strategy for Preparing Fluorogenic Tetrazine Probes with On-Demand Fluorogen Selection

Area of Science:

• Chemical Biology
• Biotechnology
• Molecular Imaging

Background:

• Fluorescent probes are crucial for life science research, linking fluorophores to functional units for biological event detection.
• Traditional probe synthesis via organic reactions requires specialized equipment and expertise, posing a barrier for life scientists.
• Developing on-demand fluorescent probes is essential for diverse experimental needs.

Purpose of the Study:

• To present a simple, rapid, and convenient strategy for preparing fluorescent probes using co-aggregation of hydrophobic molecules.
• To utilize tetrazine, a functional unit with bioorthogonal reactivity and fluorogenicity, for probe development.
• To establish a novel time-gated fluorescence bioimaging technique for multi-organelle imaging.

Main Methods:

• Co-aggregation of hydrophobic tetrazine molecules and aggregation-induced emission (AIE) luminogens in an aqueous environment.
• Quenching of AIE luminogen emission upon co-aggregation.
• Bioorthogonal click reaction to activate probe emission.
• Development of a time-gated fluorescence bioimaging method.

Main Results:

• Successfully prepared fluorogenic probes through a simple co-aggregation strategy.
• Demonstrated bioorthogonal activation of probe fluorescence via click reaction.
• Achieved multi-organelle imaging using two same-color fluorophores by distinguishing overlapping signals.
• Established a new time-gated fluorescence bioimaging technique.

Conclusions:

• The co-aggregation method offers an effective solution for on-demand fluorescent probe preparation, simplifying synthesis for life scientists.
• This approach provides valuable protocols and molecular design principles for creating novel fluorescent probes.
• The developed bioorthogonal fluorogenicity and time-gated imaging technique advance capabilities in biological research and diagnostics.