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High Throughput Zero Mode Waveguides via Natural Lithography Shawn H. Pfeil, Yale E. Goldman, NSEC DMR 0832802 Zero Mode Waveguides (ZMWs), sub-wavelength wells in a metallic substrate, can be used to create nanoscale detection volumes which allow fluorescently labeled molecules in micromolar concentrations to be interrogated one at a time 1,2 . Traditionally ZMWs have been fabricated with e-beam lithography leading to high cost and low-throughput. We are developing a high-throughput, low-cost fabrication procedure based on Natural Lithography 3 . Polystyrene beads, deposited by spin coating, form a hexagonal close-packed array. The beads are thinned using an oxygen plasma. The resulting nanoscale posts have a lattice spacing which is controlled by the original diameter of the beads (AFM images, above). Subsequent fabrication steps, such as metal deposition followed by bead lift-off, are used to create an array of wells. Fluorescently labeled proteins (left) are used to visualize an array of wells in a aluminum film. 1) Levene et al. Science 2003 2) Miyake et al. Analytical Chemistry 2008 3) Haginoya et al. Applied Physics Letters Schematic of a ZMW, modified from Miyake et al. (2008). A nanoscale observation volume is created by the evanescent electric field in a a 50-100 nm diameter well. tRN A Fluorescent N ear Field Em ission Excitation Ribosom e mRNA tRN A B tRN A Fluorescent N ear Field Em ission Excitation Ribosom e mRNA tRN A B
