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In this study, an efficient non-enzymatic glucose sensor is proposed based on a bimetal-oxide-modified carbon fiber microelectrode (CFME). The NiO-CuO/CFME nanocomposite was synthesized by consecutive electro-deposition of CuO and NiO nanoparticles on the CFME. The morphology and structure of NiCuCFME were studied with scanning electron microscopy (SEM) and energy dispersive X-ray analysis (EDX). NiCuCFME provides both a large surface area and electrocatalytic properties to improve electrochemical reactivity toward glucose oxidation. The electrocatalytic behavior of the sensing microelectrode is analyzed with cyclic voltammetry (CV) and amperometric techniques in an alkaline medium. The fabricated glucose microsensor exhibits a linear range from 1.32 μM to 570 μM, a sensitivity of 0.07 mA μM−1 cm−2, and a low limit of detection of 0.40 μM. The NiCuCFME shows long-term stability, good reproducibility, favorable repeatability, excellent selectivity, and satisfactory applicability for glucose detection which is a promising candidate for the development of non-enzymatic glucose sensors.