A novel, highly stable, selective, and sensitive non‐enzymatic glucose sensor was developed by simple and effective modification procedure. The modification of gold microelectrodes by electrochemically deposited gold nanoparticles resulted in increase of surface area up to 37 %. The nanostructured surfaces of the gold microelectrodes obtained by different modifications were studied by confocal microscopy, atomic force microscopy, and scanning electron microscopy. The gold nanoclusters exhibit great electrocatalytic properties toward glucose with a wide linear range from 0.5 to 50 mM, with a limit of detection 218 μM, and sensitivity of 185.2 mA mM−1cm−2. Moreover, the modified microelectrodes display good reproducibility, stability, and selectivity in the presence of poisoning compounds. Due to the small dimensions of gold microelectrodes and a very small volume of the sample, the microelectrodes make a contribution to miniaturisation of the system.