This work describes the electrocatalytic oxidation of glucose on CuO microspheres prepared by initially depositing copper galvanostatically from an optimized sulfate bath on glassy carbon (GC) surface and subsequently subjecting the Cu deposits to potential cycling in 0.1 M KOH. The electrochemical behaviour and electro catalytic activity of the electrode were characterized using cyclic voltametry (CV) and chronoamperometry. Scanning electron microscopy (SEM) and X-ray Photoelectron Spectroscopy (XPS) were employed to characterize the morphological and chemical oxidation states of CuO in the film. These studies shows that CuO modified GC electrodes exhibit good catalytic performance and high stability. Anodic oxidation current density was found to be 0.0496 mA cm(-2) for 1 mM of glucose. Modified electrode was validated as a sensing matrix by carrying out invitro analysis of glucose in human blood serum and in the anolyte of the microbial fuel cell (MFC). The CuO/GC electrode shows very good sensitivity (709 mA M-1 cm(-2)) and excellent linearity (R = 0.998). Our results present the scope of using CuO modified GC substrate as non-enzymatic glucose sensor for invitro analysis and as an electrocatalyst for fuel cell application. (C) 2012 The Electrochemical Society. [DOI: 10.1149/2.087204jes] All rights reserved.