Abstract Metal oxide nanoparticles prepared by pulsed laser deposition (PLD) were applied to nonenzymatic glucose detection. NiO nanoparticles with size of 3nm were deposited on glassy carbon (GC) and silicon substrates at room temperature in an oxygen atmosphere. Transmission electron microscope (TEM) image showed nanoparticles with the size of 3nm uniformly scattered on the Si(001) substrate. Unlike co-sputtering nanoparticle and carbon simultaneously, the PLD method can easily control the surface coverage of nanoparticles on the surface of substrate by deposition time. Cyclic voltammetry was performed on the samples deposited on the GC substrates for electrochemical detection of glucose. The differences between peak currents with and without glucose was used to optimize the coverage of nanoparticles on carbon electrode. The results indicated that optimal coverage of nanoparticles on carbon electrode.