A detailed study of the luminescence quantum yield, lifetimes and the number of water molecules coordinated to lanthanide luminescent probes based on biphenylyl-7-amino-4- methyl-2(1H)-quinolinon (Bi-cs124) derivatives have been carried out using a time- resolved fluorescence assay (TRFA). Bi-cs124 was conbined with different chelating agents, specifically diethylene triamine pentaacetic acid (DTPA), ethylene diamine tetraacetic acid (EDTA), ethylene glycol tetraacetic acid (EGTA), and 3,6,9,12-Tetrakis carboxymethyl)-3,6,9,12-tetraazatetradecane-1,14-dioic acid (TTHA). The results indicate that Bi-cs 124-TTHA has a longest lifetime and smallest number of coordinated water molecules because TTHA has larger number of chelating groups. The dependence of the luminescence lifetimes on the probes concentrations was investigated. The luminescence lifetime is shorter at higher probes concentrations due to self-quenching.
Images of E-coli cells labeled with one of the lanthanide luminescent probes have been obtained by TRFA excited by excimer laser radiation (351 nm) using the total internal reflection fluorescence microscopy (TIRFM). The results indicate feasibility of using lanthanide luminescent probes for time-gated luminescence microscopy.
|