An optimized immobilization procedure based on the electroreduction of aryldiazonium salt followed by covalent attachment of a cross-linked hydrogel was used to graft glucose oxidase on a carbon surface. Scanning electrochemical microscopy (SECM) and cyclic voltammetry were used to follow the construction steps of the modified electrode. By adjusting the compactness of the layer through the electrografting reaction, the penetration of the mediator through the layer can be controlled to allow the monitoring of the enzymatic activity by both cyclic voltammetry and SECM in feedback mode. The enzymatic activity of the film is finally characterized by SECM.