We prepared silver particle dimers with single Cy5 molecules localized between coupled metal particles. The silver particles with a 20 nm diameter were chemically bound with single-stranded oligonucleotides. The dimers were formed by hybridization with double-length single-stranded oligonucleotides that contained single Cy5 molecules. The image analysis revealed that the single-molecule fluorescence was enhanced 7-fold on the metal monomer and 13-fold on the metal dimer relative to the free Cy5-labeled oligonucleotide in the absence of metal. The lifetimes were shortened on the silver monomers and further shortened on the silver dimers, demonstrating the near-field interaction mechanism of fluorophore with the metal substrate. Finite-difference time-domain (FDTD) calculations were employed to study the distribution of electric field near the metal monomer and dimer. The coupling effect of metal particle on the fluorescence enhancement was discussed.