Preparation of visible-light-excited europium biolabels for time-resolved luminescence cell imaging application.

By using a visible-light-excited ternary Eu(3+) complex, BHHBCB-Eu(3+)-BPT (BHHBCB: 1,2-bis[4'-(1",1",1",2",2",3",3"-heptafluoro-4″,6″-hexanedion-6″-yl)-benzyl]-4-chlorosulfobenzene; BPT: 2-(N,N-diethylanilin-4-yl)-4,6-bis(pyrazol-1-yl)-1,3,5-triazine), as a luminophore, two kinds of novel visible-light-excited europium materials, the silica-encapsulated BHHBCB-Eu(3+)-BPT (Eu@SiO2) nanoparticles and BHHBCB-Eu(3+)-BPT-conjugated bovine serum albumin (BSA-BHHBCB-Eu(3+)-BPT), were prepared for biolabeling and time-resolved luminescence cell imaging applications. The Eu@SiO2 nanoparticles, prepared by the copolymerization of 3-aminopropyl(triethoxy)silane-BHHBCB-Eu(3+)-BPT conjugate, free 3-aminopropyl(triethoxy) silane and tetraethyl orthosilicate in a water-in-oil reverse microemulsion, are monodispersed, spherical and uniform in size, and strongly luminescent with an excitation peak at ≈ 400 nm and a long luminescence lifetime of 346 µs. The BSA-BHHBCB-Eu(3+)-BPT, prepared by covalent binding of BHHBCB-Eu(3+)-BPT to BSA, shows also strong visible-light-excited luminescence with a excitation peak at ≈ 400 nm and a long luminescence lifetime of 402µs. The two materials were used for labeling transferrin and folic acid. Using the time-resolved luminescence imaging of living HeLa cells, the cell-surface receptors of transferrin and folic acid were successfully visualized by the prepared biolabels based on the ligand-receptor affinity binding interaction. The results demonstrated the feasibility of the new materials as visible-light-excited biolabels for the time-resolved luminescence cell imaging.