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The plasmonic effects of metallic nanostructures in fluorescence have been receiving considerable attention because of their promises in new methods,probes and devices for the utilization of fluorescence in many important applications,such as bioanalysis,biomedical research,and clinical testing1.Here we described the dequenching of fluorescence in surface plasmon-coupled emission (SPCE)2 through a sphere-on-surface geometry3,4,where the surface-bound silver nanoparticles dequenched the signal of fluorophores coated on gold nano-film,radiating the directional p-polarized emission through the adhered prism at a defined angle.We discovered that the special dequenched fluorescence originating from the coupling of fluorophores with localized surface plasmons on nanoparticles and propagating surface plasmons on nano-film,possessed unique characteristics suitable and beneficial to biosensing design.As illustrated in Figure 1,a model immunoassay is established based on the dequenching behavior,with numerous preloaded labels revived to sensitively report the biorecognition,which demonstrates its potential to create a new generation of plasmon-assisted fluorescence biosensing.