| Bioorthogonal reactions, characterized by high yield, exceptional selectivity, efficiency, and minimal side reactions, have been extensively employed for labeling biomolecules in living systems, targeted imaging, and the visualization of biological processes. Tetrazine-boron dipyrromethene (BODIPY) derivatives serve as exemplary probes whose fluorescence modulation is governed by a distinctive mechanism: upon conjugation, the tetrazine moiety effectively quenches BODIPY fluorescence through multiple pathways, including photoinduced electron transfer (PET), Förster resonance energy transfer (FRET), and through-bond energy transfer (TBET). Subsequent reaction with a dienophile via an inverse electron demand Diels–Alder (IEDDA) cycloaddition restores the extended conjugation and reduces the energy gap, leading to a pronounced recovery of fluorescence. In recent years, BODIPY–tetrazine derivatives have achieved significant breakthroughs in live-cell tracking, tumor microenvironment imaging, and the labeling of proteins and nucleic acids. Their rapid responsiveness, high signal-to-noise ratio, and robust in vivo stability have established them as cutting-edge tools in biofluorescence imaging. |
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