Cell assay using a two-photon-excited europium chelate.

We report application of two-photon excitation of europium chelates to immunolabeling of epidermal growth factor receptor (EGFR) cell surface proteins on A431 cancer cells. The europium chelates are excited with two photons of infrared light and emit in the visible. Europium chelates are conjugated to antibodies for EGFR.

Strategy for photostable proximity bioassays using lanthanides.

We report initial findings for research aimed at creating photostable lanthanide chelate reporters for proximity assays. These reporters take advantage of the nanometer-scale distance dependence of fluorescence enhancement for molecules in the vicinity of noble metal nanoparticles and also capitalize on some unique properties of lanthanide chelates. This approach promises to lead to proximity assays that do not suffer from photobleaching and offer very high on/off enhancement ratios.

Upconversion from aqueous phase lanthanide chelates.

We have prepared and characterized several lanthanide ion complexes of multidentate ligands or chelates in an effort to develop new upconverting luminescent labels that can be immune to autofluorescence and photobleaching. This study has involved the characterization of various chelates of Nd, Er, and Tm with respect to relative luminescent efficiency and excited-state lifetimes and explored various two-photon stepwise excitation mechanisms. Using peak laser powers near 100 kW, the upconversion emissions of Nd in Nd(EDTA)2(5-) at 386 nm, Er in Er(DPA)3(3-) at 550 nm, and Tm in Tm(DPA)3(3-) at 480 nm, at levels of approximately 10(-12) moles can be detected.