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Modern biomedical research and preclinical pharmaceutical development rely heavily on the phenotyping of small vertebrate models for various diseases prior to human testing. In this article, we demonstrate an acoustofluidic rotational tweezing platform that enables contactless, high-speed, 3D multispectral imaging and digital reconstruction of zebrafish larvae for quantitative phenotypic analysis. The acoustic-induced polarized vortex streaming achieves contactless and rapid (~1 s/rotation) rotation of zebrafish larvae. This enables multispectral imaging of the zebrafish body and internal organs from different viewing perspectives. Moreover, we develop a 3D reconstruction pipeline that yields accurate 3D models based on the multi-view images for quantitative evaluation of basic morphological characteristics and advanced combinations of metrics. With its contactless nature and advantages in speed and automation, our acoustofluidic rotational tweezing system has the potential to be a valuable asset in numerous fields, especially for developmental biology, small molecule screening in biochemistry, and pre-clinical drug development in pharmacology.

 

Contact-free manipulation of small objects ( e.g. , cells, tissues, and droplets) using acoustic waves eliminates physical contact with structures and undesired surface adsorption. Pioneering acoustic-based, contact-free manipulation techniques ( e.g. , acoustic levitation) enable programmable manipulation but are limited by evaporation, bulky transducers, and inefficient acoustic coupling in air. Herein, we report an acoustofluidic mechanism for the contactless manipulation of small objects on water. A hollow-square-shaped interdigital transducer (IDT) is fabricated on lithium niobate (LiNbO 3 ), immersed in water and used as a sound source to generate acoustic waves and as a micropump to pump fluid in the ± x and ± y orthogonal directions. As a result, objects which float adjacent to the excited IDT can be pushed unidirectionally (horizontally) in ± x and ± y following the directed acoustic wave propagation. A fluidic processor was developed by patterning IDT units in a 6-by-6 array. We demonstrate contactless, programmable manipulation on water of oil droplets and zebrafish larvae. This acoustofluidic-based manipulation opens avenues for the contactless, programmable processing of materials and small biosamples.