Zero-mode waveguides (ZMWs) are optical nanostructures to confine fluorescent excitation within sub-diffraction
volumes and are commonly used for single-molecule analysis. However, the conventional ZMWs with aluminum film on
fused silica have limitations on living cell studies. The same surfaces composed of hydroxyl group inside and outside
each ZMW restrict specific surface functionalization. The sharp-cylinder shaped and rough edge of each ZMW produces
a steric interference of molecular dynamics on cell membrane. In this study, selectively surface functionalization inside
and outside of each ZMW was achieved with tri-metal-layer film on fused silica. Moreover, bowl-shaped and smooth edge of each ZMW was manufactured in large area. The improvement of ZMWs provides a broad way for monitoring molecular dynamics in living cells.
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