To investigate how nano-aggregation and AIEgens will affect the turn-on emission of tetrazine probes, we designed and synthesized a series of tetrazine-dye conjugates. Tetrazine units were synthesized under Pinner synthesis using thiol as a catalyst, and sequential Pd-catalyzed coupling reactions connected tetrazine to chromophores. The obtained compounds were recrystallized, and purity was confirmed by High-Performance Liquid Chromatography (HPLC) to avoid impurity effects (shown in Figure -S90, Supporting Information). Their molecular structures were thoroughly characterized by nuclear magnetic resonance (NMR) spectroscopy and high-resolution mass spectra (shown in Figures , Supporting Information). Single crystals were obtained except for MTPA-Tz, and X-ray diffraction analysis supports the structure (Figures , in Supporting Information). We prepared four AIEgen-tetrazine conjugates (AIE-Tz) and one normal planar fluorophore-tetrazine conjugate (PR-Tz) as a negative control for the entire experiment. To obtain a multifaceted and general perspective of this strategy, we designed AIEgens with diverse structures that include either tetraphenyl ethylene (TPE-Tz) or triaryl amines (TPA-Tz, MTPA-Tz, and MBTPA-Tz) as well-known twisted units to achieve AIE properties. As expected, all molecules reacted quantitatively with the strained alkyne compound (1R,8S,9s)-Bicyclo[6.1.0]non-4yn-9-ylmethanol (BCN-OH) to provide emissive pyridazine compounds (Figure ), and the obtained AIE-Pzs covered a broad emission range from violet to NIR (380 to 780 nm) in water (Figure ) with an emission maximum at 419 nm, 481 nm, 538 nm, 636 nm for TPA-Pz, TPA-Pz, MTPA-Pz, and MBTPA-Pz, respectively.