Fig. 1

a Flow chart of the working mechanism of the ART system. b Illustration of the experimental configuration of the acoustofluidic chip for rotational manipulation of zebrafish larvae mounted on a conventional optical microscope platform. The chip consists of an IDT fabricated on a LiNbO₃ piezoelectric substrate which generates acoustic waves and a patterned fluidic channel aligned parallel to the lateral side of the IDT (y axis) with half of its width on the IDT. The zebrafish larvae in the channel can be rotated by polarized acoustic streaming in a single vortex pattern in the y–z planes, which was induced by acoustic waves propagating in the ±x direction. The three key parameters contributing to the features of the vortex tube are denoted by “a” for the width of the square cross-section of the channel, “e” for the width of the effective IDT area, and “L” for the length of the IDT. c Multiple labeled organs of the larvae can be imaged using the corresponding fluorescent wavelength during rotation. Scale bar: 1 mm. d From this multi-angle sequence of microscope images, 3D models of different internal organs of interest can be reconstructed, assembled, and quantified as digital readouts using a computer-vision-based algorithm for subsequent quantitative phenotypic analysis of morphological characteristics.