Localized surface plasmon resonance (LSPR) excited by an oblique incidence of S- and P-polarized light to a two-dimensionally assembled silver nanoparticle sheet was investigated via enhanced fluorescence under total internal reflection fluorescence (TIRF) microscopy. The finite-difference-time-domain simulation demonstrated that the S-polarized light induced a strong plasmon coupling at a nanogap between the particles, which eventually led to a highly confined, strong, and “flattened” electric field on the entire surface. In contrast, the LSPR field excited by P-polarized light was located on the individual particles, having a relatively long tail in the axial direction (low confinement). The LSPR-mediated fluorescence appeared stronger under P-polarized light than under S-polarized light in the experiments using cyanine dye solutions, while the opposite result was obtained for the fluorescence bead snapshot (diameter: 200 nm). Magnified images of the single beads taken by a super-resolution digital CMOS camera (65 nm/pixel) revealed improved lateral resolution when S-polarized light was used on both the silver nanoparticle sheet and glass under TIRF microscopy.

This paper reports our original technique for visualizing cell-attached nanointerfaces with extremely high axial resolution using homogeneously excited localized surface plasmon resonance (LSPR) on self-assembled silver nanoparticle sheets. The LSPR sheet can confine and enhance the fluorescence at the nanointerface, which provides high signal-to-noise ratio images of focal adhesion at the cell-attached interface. The advantage of this LSPR-assisted technique is its usability, which provides comparable or higher-quality nanointerfacial images than TIRF microscopy, even under epifluorescence microscopy. We also report the cytotoxicity of silver nanoparticles, as determined via morphological analysis of adherent cells on the sheet.

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