Acoustic levitation and manipulation of columns of droplets with integrated optical detection for parallelisation of reactions

The most common methodology for performing multiple chemical and biological reactions in parallel is to use microtitre plates with either manual or robotic dispensing of reactants and wash solutions. We envision a paradigm shift where acoustically levitated droplets serve as wells of microtitre plates and are acoustically manipulated to perform chemical and biological reactions in a non-contact fashion. This in turn requires that lines of droplets can be levitated and manipulated simultaneously so that the same operations (merge, mix, and detect) can be performed on them in parallel. However, this has not been demonstrated until this work. Because of the nature of acoustic standing waves, a single focus has more than one trap, and can allow levitation of columns of droplets at the focal point and at half a wavelength above and below that point. Using this approach, we increased the number of acoustically levitated and merged droplets to 6 compared to 2 in the state-of-the-art. We showed that droplets in a column can be moved and merged with droplets in another column simultaneously and in a controlled manner to perform repeats and/or parallelisation of chemical and biological reactions. To demonstrate our approach experimentally, we built an acoustic levitator with top and bottom surfaces made of a 16 x 16 grid of 40 kHz phased array transducers and integrated optical detection system, studied two acoustic trap generation and movement algorithms, and performed an exemplar enzyme assay. This work has made significant steps towards acoustic levitation and manipulation of large numbers of droplets to eventually significantly reduce the use of the current state-of-the-art tools, microtitre plates and robots, for performing parallelised chemical and biological reactions. In this work, acoustically levitated droplets serve as wells of microtitre plates and are acoustically manipulated to perform chemical and biological reactions in a non-contact fashion.