<p>Rotational manipulation of microscale objects or living organisms is crucial for various applications in microsystems and biological engineering, especially in high-precision imaging. However, achieving precise rotation of individual biological samples at specific angles remains a challenge. This study introduces a novel and cost-effective acoustofluidic approach for on-demand rotation of microparticles and biological specimens, such as zebrafish larvae, to desired angles. The system consists of piezoelectric transducers arranged orthogonally on a structured glass capillary. By applying sinusoidal signals with a defined phase difference (<i>φ</i>) to these transducers, we can adjust the ultrasonic resonant field in the capillary’s microchannel to rotate the acoustic pressure nodal plane at the cross-section. This rotation creates an acoustic radiation force that traps and aligns suspended objects along the rotated pressure node. Within a specific operational range, specimens are securely trapped along the channel axis and rotated at a controlled angle <i>α</i> relative to <i>φ</i> (following the relationship <i>α</i> ≈ <i>φ</i>/2), demonstrating stepwise rotational control. By dynamically adjusting the phase difference <i>φ</i> from 0 to 4<i>π</i>, we can achieve controlled full 360° rotation of a trapped object. Further parametric numerical studies are conducted to showcase the effectiveness of this technique at various operating parameters for the controlled rotational manipulation of objects suspended in a fluid. The use of a glass capillary and external piezoelectric transducers offers a smooth and biologically permissive environment, simplifying fabrication and showing significant potential for applications in micro-robotics, single-cell analysis, and organism-level studies in fields like developmental biology.</p><p></p>