Reduced acoustic resonator dimensions improve focusing efficiency of bacteria and submicron particles

In this study, we demonstrate an acoustofluidic device that enables single-file focusing of submicron particles and bacteria using a two-dimensional (2D) acoustic standing wave. The device consists of a 100 mu m x 100 mu m square channel that supports 2D particle focusing in the channel center at an actuation frequency of 7.39 MHz. This higher actuation frequency compared with conventional bulk acoustic systems enables radiation-force-dominant motion of submicron particles and overcomes the classical size limitation (approximate to 2 mu m) of acoustic focusing. We present acoustic radiation force-based focusing of particles with diameters less than 0.5 mu m at a flow rate of 12 mu L min(-1), and 1.33 mu m particles at flow rates up to 80 mu L min(-1). The device focused 0.25 mu m particles by the 2D acoustic radiation force while undergoing a channel cross-section centered, single-vortex acoustic streaming. A suspension of bacteria was also investigated to evaluate the biological relevance of the device, which demonstrated the alignment of bacteria in the channel at a flow rate of up to 20 mu L min(-1). The developed acoustofluidic device can align submicron particles within a narrow flow stream in a highly robust manner, validating its use as a flow-through focusing chamber to perform high-throughput and accurate flow cytometry of submicron objects.