The nanosphere lithography protocol presented here for fabricating zero mode waveguides is an accessible, low cost method that does not require any specialized fabrication tools or facilities. This method will be useful for single molecule biophys
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Described here is a nanosphere lithography method for parallel fabrication of zero mode waveguides, which are arrays of nanoapertures in a metal-clad glass microscopy coverslip for single molecule imaging at nano- to micromolar concentrations of fluorophores. The method takes advantage of colloidal crystal self-assembly to create a waveguide template.