Homogeneous and fast mixing of samples at microscale is a critical requirement for successful applications of microfluidics in biochemical analysis, chemical synthesis, drug delivery and nanomaterial synthesis. This paper reports the optimisation of bubble-induced mixing in a microfluidic device in terms of voltage, driving frequency, piezo transducer position and PDMS thickness. The microfluidic device consists of a microwell (with the diameter of 1mm and volume of ~95 nL) with two rectangular bubble traps (400×400μm) on both sides of the well. After the injection of liquid, air bubbles were spontaneously trapped in two rectangular traps. When the frequency of a piezo was equal to the resonance frequency of air bubbles, strong liquid recirculation formed (so called acoustic microstreaming) in the vicinity of the interface of air bubbles and water. The acoustic induced flow of microbeads and mixing of water and fluorescence dye were imaged to study the mixing efficiency. For a given voltage and PDMS thickness, when the piezo was placed on top of the well, the mixing was most vigorous. For a given frequency, the mixing efficiency was directly proportional to the voltage (4-20V) and inversely proportional to the PDMS thickness (0.3-2mm). When the frequency driving the piezo was approaching the resonance frequency of air bubbles, the mixing efficiency was maximal, while when it was far away from the resonance frequency of air bubbles, the mixing efficiency was much lower. This work provides guidance to the design and the application of bubble-induced acoustic mixing in microfluidics.