Acoustic standing wave technology combined with microtechnology opens up new areas for the development of advanced particle and cell separating microfluidic systems. This tutorial review outlines the fundamental work performed on continuous flow acoustic standing wave separation of particles in macro scale systems. The transition to the microchip format is further surveyed, where both fabrication and design issues are discussed. The acoustic technology offers attractive features, such as reasonable throughput and ability to separate particles in a size domain of about tenths of micrometers to tens of micrometers. Examples of different particle separation modes enabled in microfluidic chips, utilizing standing wave technology, are described along a discussion of several potential applications in life science research and in the medical clinic. Chip integrated acoustic standing wave separation technology is still in its infancy and it can be anticipated that new laboratory standards very well may emerge from the current research.

The progress in microfabrication and lab-on-a-chip technologies has been a major area of development for new approaches to bioanalytics and integrated concepts for cell biology. Fundamental advances in the development of elastomer based microfluidics have been driving factors for making microfluidic technology available to a larger scientific community in the past years. In line with this, microfluidic separation of cells and particles is currently developing rapidly where key areas of interest are found in designing lab-on-a-chip systems that offer controlled microenvironments for studies of fundamental cell biology. More recently industrial interests are seen in the development of micro chip based flow cytometry technology both for preclinical research and clinical diagnostics. This critical review outlines the most recent developments in microfluidic technology for cell and particle separation in continuous flow based systems. (130 references)

This acoustofluidics tutorial focuses on continuous flow-based half wavelength resonator systems operated in the transversal mode, where the direction of the primary acoustic force acts in plane with the microchip. The transversal actuation mode facilitates integration with up- and downstream microchannel networks as well as visual control of the acoustic focusing experiment. Applications of particle enrichment in an acoustic half wavelength resonator are discussed as well as clarification of the carrier fluid from undesired particles. Binary separation of particle/vesicle/cell mixtures into two subpopulations is outlined based on the different polarities of the acoustic contrast factor. Furthermore, continuous flow separation of different particle/cell types is described where both Free Flow Acoustophoresis (FFA) and binary acoustophoresis are utilized. By capitalizing on the laminar flow regime, acoustophoresis has proven especially successful in performing bead/cell translations between different buffer systems. Likewise, the ability to controllably translate particulate matter across streamlines has opened a route to valving of cells/particles without any moving parts, where event triggered cell sorting is becoming an increasing area of activity. Recent developments now also enable measurements of fundamental cell properties such as density and compressibility by means of acoustophoresis. General aspects on working with live cells in acoustophoresis systems are discussed as well as available means to quantify the outcome of cell and particle separation experiments performed by acoustophoresis.