A three-dimensionally ordered, macroporous, inverse-opal platinum film was synthesized electrochemically by the inverted colloidal-crystal template technique. The inverse-opal film that contains platinum nanoparticles showed improved electrocatalytic activity toward glucose oxidation with respect to the directly deposited platinum; this improvement is due to the interconnected porous structure and the greatly enhanced effective surface area. In addition, the inverse-opal Pt-film electrode responds more sensitively to glucose than to common interfering species of ascorbic acid, uric acid, and p-acetamidophenol due to their different electrochemical reaction mechanisms. Results showed that the ordered macroporous materials with enhanced selectivity and sensitivity are promising for fabrication of nonenzymatic glucose biosensors.