Nonlinear optical detection of proteins based on localized surface plasmons in surface immobilized gold nanospheres

A new nonlinear optical method is presented to detect proteins binding to a gold surface without using fluorescent-dye labeling. After exposure of the protein-binding surface to a gold nanosphere solution, the nanospheres are immobilized above a gold surface with a nanogap supported by the protein. The gold nanospheres immobilized on the gold surface show strong localized surface plasmon (LSP) resonance, and the formation of this structure results in a marked increase in the optical second harmonic (SH) activity of the gold surface arising from a large enhancement of the electric field localized adjacent to the nanospheres on the LSP resonance. The SH image, therefore, gives a high contrast ratio, 7.0:1, of protein-binding spots to control spots. The contrast ratio is much greater than those obtained by linear reflectivity imaging.