Significance: There is a lack of a general method to noninvasively and remotely manipulate cells with high spatiotemporal precisions. We developed an ultrasound-based mechanogenetics system to achieve this goal. Cells were engineered with the mechanosensor Piezo1 and genetic transducing modules to perceive the mechanical perturbation generated by the ultrasound wave and transduce it into genetic activities. Mechanosensitive and ultrasound-controllable T cells were further engineered to target and eradicate tumor cells with inducible chimeric antigen receptors. This mechanogenetics approach can be extended to remotely control, in principle, any gene activity in live cells for the reprogramming of cellular functions. This method should also provide a general approach to remotely control molecular functions for biological studies and clinical applications, particularly cell-based cancer immunotherapy.

High frequency ultrasonic imaging is considered by many to be the next frontier in ultrasound. It has many clinical applications ranging from imaging the eye and skin to small animal imaging. Small animal imaging has recently generated intense interest for the purpose of evaluating the efficacy of drugs and gene therapy. Commercial high frequency scanners often termed “ultrasonic biomicroscope”, or UBM, all use mechanically scanned single element transducers at frequencies between 30 to 60 MHz with a frame rate of 30 frames/second or lower. To alleviate problems with UBMs which include mechanical motion and fixed focusing, high frequency linear arrays and imaging systems in the 20–50 MHz range have been developed. In this paper, current efforts in the development of high frequency ultrasonic imaging will be reviewed and potential biomedical applications discussed.

Purpose: Conventional machining methods for fabricating piezoelectric components such as ultrasound transducer arrays are time-consuming and limited to relatively simple geometries. The purpose of this paper is to develop an additive manufacturing process based on the projection-based stereolithography process for the fabrication of functional piezoelectric devices including ultrasound transducers. Design/methodology/approach: To overcome the challenges in fabricating viscous and low-photosensitive piezocomposite slurry, the authors developed a projection-based stereolithography process by integrating slurry tape-casting and a sliding motion design. Both green-part fabrication and post-processing processes were studied. A prototype system based on the new manufacturing process was developed for the fabrication of green-parts with complex shapes and small features. The challenges in the sintering process to achieve desired functionality were also discussed. Findings: The presented additive manufacturing process can achieve relatively dense piezoelectric components (approximately 95 per cent). The related property testing results, including X-ray diffraction, scanning electron microscope, dielectric and ferroelectric properties as well as pulse-echo testing, show that the fabricated piezo-components have good potentials to be used in ultrasound transducers and other sensors/actuators. Originality/value: A novel bottom-up projection system integrated with tape casting is presented to address the challenges in the piezo-composite fabrication, including small curing depth and viscous ceramic slurry recoating. Compared with other additive manufacturing processes, this method can achieve a thin recoating layer (as small as 10 μm) of piezo-composite slurry and can fabricate green parts using slurries with significantly higher solid loadings. After post processing, the fabricated piezoelectric components become dense and functional.

High-frequency ultrasonic imaging with improved spatial resolution has gained increasing attention in the field of biomedical imaging. Sensitivity of transducers plays a pivotal role in determining ultrasonic image quality. Conventional ultrasonic transducers are mostly made from lead-based piezoelectric materials that may be harmful to the human body and the environment. In this study, a new (K,Na)NbO $_{3}$ -KTiNbO $_5$ -BaZrO $_{3}$ -Fe $_{2}$ O $_{3}$ -MgO (KNN-NTK-FM) lead-free piezoelectric ceramic was utilized in developing eco-friendly transducers for high-frequency biomedical ultrasonic imaging applications. A needle transducer with a small active aperture size of 0.45 × 0.55 mm $^2$ was designed and evaluated. The fabricated transducer exhibits great performance with a high center frequency (52.6 MHz), a good electromechanical coupling ( $k$ $_{eff}$ ~ 0.45), a large bandwidth (64.4% at -6 dB), and a very low two-way insertion loss (10.1 dB). Such high sensitivity is superior to those transducers based on other lead-free piezoelectric materials and can even be comparable to the lead-based ones. Imaging performance of the KNN-NTK-FM needle transducer was analyzed by imaging a wire phantom and an agar tissue-mimicking phantom. Imaging capabilities of the transducer were further demonstrated by ex vivo imaging studies on a porcine eyeball and a rabbit aorta. The results suggest that the KNN-NTK-FM piezoceramic has many attractive properties over other lead-free piezoelectric materials in developing eco-friendly highly sensitive transducers for high-frequency biomedical ultrasonic imaging applications.

We report on a real-time acoustic radiation force optical coherence elastography (ARF-OCE) system to map the relative elasticity of corneal tissue. A modulated ARF is used as excitation to vibrate the cornea, while OCE serves as detection of tissue response. To show feasibility of detecting mechanical contrast using this method, we performed tissue-equivalent agarose phantom studies with inclusions of a different stiffness. We obtained 3-D elastograms of a healthy cornea and a highly cross-linked cornea. Finally, we induced a stiffness change on a small portion of a cornea and observed the differences in displacement.

Controlling cell functions for research and therapeutic purposes may open new strategies for the treatment of many diseases. An efficient and safe introduction of membrane impermeable molecules into target cells will provide versatile means to modulate cell fate. We introduce a new transfection technique that utilizes high frequency ultrasound without any contrast agents such as microbubbles, bringing a single-cell level targeting and size-dependent intracellular delivery of macromolecules. The transfection apparatus consists of an ultrasonic transducer with the center frequency of over 150 MHz and an epi-fluorescence microscope, entitled acoustic-transfection system. Acoustic pulses, emitted from an ultrasonic transducer, perturb the lipid bilayer of the cell membrane of a targeted single-cell to induce intracellular delivery of exogenous molecules. Simultaneous live cell imaging using HeLa cells to investigate the intracellular concentration of Ca²⁺ and propidium iodide (PI) and the delivery of 3 kDa dextran labeled with Alexa 488 were demonstrated. Cytosolic delivery of 3 kDa dextran induced via acoustic-transfection was manifested by diffused fluorescence throughout whole cells. Short-term (6 hr) cell viability test and long-term (40 hr) cell tracking confirmed that the proposed approach has low cell cytotoxicity.