Robust discrimination between single gold nanoparticles and their dimers in aqueous solution for ultrasensitive homogeneous bioassays

We propose a robust method to distinguish isolated single gold nanoparticles (AuNP monomers) and their dimers under Brownian motion, a key for ultrasensitive homogeneous bioassays, including AuNP sandwich assays. To detect dimers and distinguish them from a larger number of monomers in aqueous solution, single-particle polarization microscopy was performed. For the accurate detection of individual particles, the optical anisotropy and rotational diffusion time are measured because a dimer is much more anisotropic than the nearly spherical monomer and the rotational diffusion time of a dimer is four times that of a monomer. By employing an autocorrelation analysis, we defined a measure of distinguishing that simultaneously enables high detection probability and low error probability. The detection platform offers homogeneous DNA hybridization assays and immunoassays at the subpicomolar level.