A simple approach for in situ laser-induced modification of iridium-based materials to increase their electrocatalytic activity towards enzyme-free glucose sensing was proposed. For this purpose, we deposited gold and platinum separately and as a mixture on the surface of pre-synthesized iridium microstructures upon laser irradiation at a wavelength of 532 nm. Then, we carried out the comparative investigation of their morphology, elemental and phase composition as well as their electrochemical properties. The best morphology and, as a result, the highest sensitivity (~9960 µA/mM cm2) with respect to non-enzymatic determination of D-glucose were demonstrated by iridium-gold-platinum microstructures also showing low limit of detection (~0.12 µM), a wide linear range (0.5 µM–1 mM) along with good selectivity, reproducibility and stability.