Fabrication and Characterization of a Miniaturized 15-MHz Side-Looking Phased-Array Transducer Catheter.

This paper describes the development of a miniaturized 15-MHz side-looking phased-array transducer catheter. The array features a 2-2 linear composite with 64 piezoelectric elements mechanically diced into a piece of PMN-30%PT single crystal and separated by non-conductive epoxy kerfs at a 50-µm pitch, yielding a total active aperture of 3.2 mm in the azimuth direction and 1.8 mm in the elevation direction, with an elevation natural focal depth of 8.1 mm. The array includes non-conductive epoxy backing and two front matching layers. A custom flexible circuit connects the array piezoelectric elements to a bundle of 64 individual 48-AWG micro-coaxial cables enclosed within a 1.5-m long 10F catheter. Performance characterization was evaluated via finite element analysis simulations and afterwards compared against obtained measurement results, which showed an average center frequency of 17.7 MHz, an average bandwidth of 52.2% at -6 dB, and crosstalk less than -30 dB. Imaging of a tungsten fine-wire phantom resulted in axial and lateral spatial resolutions of approximately 90 µm and 420 ìm, respectively. The imaging capability was further evaluated with colorectal tissue-mimicking phantoms, demonstrating the potential suitability of the proposed phased-array transducer for the intraoperative assessment of surgical margins during minimally invasive colorectal surgery procedures.

Investigation of Optimized Treatment Conditions for Acoustic-Transfection Technique for Intracellular Delivery of Macromolecules.

Manipulation of cellular functions and structures by introduction of genetic materials inside cells has been one of the most prominent research areas in biomedicine. High-frequency ultrasound acoustic-transfection has recently been developed and confirmed by intracellular delivery of small molecules into HeLa cells at the single-cell level with high cell viability. After we proved the concept underlying the acoustic-transfection technique, treatment conditions for different human cancer cell lines have been intensively investigated to further develop acoustic-transfection as a versatile and adaptable transfection method by satisfying the requirements of high-delivery efficiency and cell membrane permeability with minimal membrane disruption. To determine optimal treatment conditions for different cell lines, we developed a quantitative intracellular delivery score based on delivery efficiency, cell membrane permeability and cell viability after 4 and 20 h of treatment. The intracellular delivery of macromolecules and the simultaneous intracellular delivery of two molecules under optimal treatment conditions were successfully achieved. We found that DNA plasmid was delivered by acoustic-transfection technique into epiblast stem cells, which expressed transient mCherry fluorescence.

Fabrication and Characterization of a 20-MHz Microlinear Phased-Array Transducer for Intervention Guidance.

This paper describes the design and fabrication of a miniature ultrasonic phased-array transducer used for intervention guidance. Currently, ultrasound probes are often placed at the body surface of the patients, leading to several drawbacks including the limitation of penetration and image quality. In order to improve the reliability of the guiding process, we propose a miniature phased-array transducer that can be placed adjacent to the intervention device during the interventional procedure. In this paper, we report the work that has been carried out on the development of this miniature phased-array transducer. It comprised 48 elements housed in a 3-mm-diameter needle. A specially designed flexible circuit was used for accommodating the transducer array in the long, thin needle housing. The center frequency and the fractional bandwidth were approximately 20 MHz and 42%, respectively, with an average crosstalk lower than -30 dB. The axial and azimuth resolutions were approximately 80 and [Formula: see text], respectively. The imaging capability of the transducer was further evaluated by acquiring the B-mode images of a needle in a cow liver. The performance of the proposed phased-array transducer demonstrates the feasibility of such an approach for interventional guidance.

Contactless microparticle control via ultrahigh frequency needle type single beam acoustic tweezers.

This paper reports on contactless microparticle manipulation including single-particle controlled trapping, transportation, and patterning via single beam acoustic radiation forces. As the core component of single beam acoustic tweezers, a needle type ultrasonic transducer was designed and fabricated with center frequency higher than 300 MHz and -6 dB fractional bandwidth as large as 64%. The transducer was built for an f-number close to 1.0, and the desired focal depth was achieved by press-focusing technology. Its lateral resolution was measured to be better than 6.7 µm by scanning a 4 µm tungsten wire target. Tightly focused acoustic beam produced by the transducer was shown to be capable of manipulating individual microspheres as small as 3 µm. "USC" patterning with 15 µm microspheres was demonstrated without affecting nearby microspheres. These promising results may expand the applications in biomedical and biophysical research of single beam acoustic tweezers.

Ultrahigh Frequency (100 MHz-300 MHz) Ultrasonic Transducers for Optical Resolution Medical Imagining.

High resolution ultrasonic imaging requires high frequency wide band ultrasonic transducers, which produce short pulses and highly focused beam. However, currently the frequency of ultrasonic transducers is limited to below 100 MHz, mainly because of the challenge in precise control of fabrication parameters. This paper reports the design, fabrication, and characterization of sensitive broadband lithium niobate (LiNbO3) single element ultrasonic transducers in the range of 100-300 MHz, as well as their applications in high resolution imaging. All transducers were built for an f-number close to 1.0, which was achieved by press-focusing the piezoelectric layer into a spherical curvature. Characterization results demonstrated their high sensitivity and a -6 dB bandwidth greater than 40%. Resolutions better than 6.4 µm in the lateral direction and 6.2 µm in the axial direction were achieved by scanning a 4 µm tungsten wire target. Ultrasonic biomicroscopy images of zebrafish eyes were obtained with these transducers which demonstrate the feasibility of high resolution imaging with a performance comparable to optical resolution.

Direct and sustained intracellular delivery of exogenous molecules using acoustic-transfection with high frequency ultrasound.

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 Ca(2+) 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.

Design of matching layers for high-frequency ultrasonic transducers.

Matching the acoustic impedance of high-frequency (≥100 MHz) ultrasound transducers to an aqueous loading medium remains a challenge for fabricating high-frequency transducers. The traditional matching layer design has been problematic to establish high matching performance given requirements on both specific acoustic impedance and precise thickness. Based on both mass-spring scheme and microwave matching network analysis, we interfaced metal-polymer layers for the matching effects. Both methods hold promises for guiding the metal-polymer matching layer design. A 100 MHz LiNbO3 transducer was fabricated to validate the performance of the both matching layer designs. In the pulse-echo experiment, the transducer echo amplitude increased by 84.4% and its -6dB bandwidth increased from 30.2% to 58.3% comparing to the non-matched condition, demonstrating that the matching layer design method is effective for developing high-frequency ultrasonic transducers.

High-resolution harmonic motion imaging (HR-HMI) for tissue biomechanical property characterization.

BACKGROUND: Elastography, capable of mapping the biomechanical properties of biological tissues, serves as a useful technique for clinicians to perform disease diagnosis and determine stages of many diseases. Many acoustic radiation force (ARF) based elastography, including acoustic radiation force impulse (ARFI) imaging and harmonic motion imaging (HMI), have been developed to remotely assess the elastic properties of tissues. However, due to the lower operating frequencies of these approaches, their spatial resolutions are insufficient for revealing stiffness distribution on small scale applications, such as cancerous tumor margin detection, atherosclerotic plaque composition analysis and ophthalmologic tissue characterization. Though recently developed ARF-based optical coherence elastography (OCE) methods open a new window for the high resolution elastography, shallow imaging depths significantly limit their usefulness in clinics. METHODS: The aim of this study is to develop a high-resolution HMI method to assess the tissue biomechanical properties with acceptable field of view (FOV) using a 4 MHz ring transducer for efficient excitation and a 40 MHz needle transducer for accurate detection. Under precise alignment of two confocal transducers, the high-resolution HMI system has a lateral resolution of 314 µm and an axial resolution of 147 µm with an effective FOV of 2 mm in depth. RESULTS: The performance of this high resolution imaging system was validated on the agar-based tissue mimicking phantoms with different stiffness distributions. These data demonstrated the imaging system's improved resolution and sensitivity on differentiating materials with varying stiffness. In addition, ex vivo imaging of a human atherosclerosis coronary artery demonstrated the capability of high resolution HMI in identifying layer-specific structures and characterizing atherosclerotic plaques based on their stiffness differences. CONCLUSIONS: All together high resolution HMI appears to be a promising ultrasound-only technology for characterizing tissue biomechanical properties at the microstructural level to improve the image-based diseases diagnosis in multiple clinical applications.

Non-contact high-frequency ultrasound microbeam stimulation for studying mechanotransduction in human umbilical vein endothelial cells.

We describe how contactless high-frequency ultrasound microbeam stimulation (HFUMS) is capable of eliciting cytoplasmic calcium (Ca(2+)) elevation in human umbilical vein endothelial cells. The cellular mechanotransduction process, which includes cell sensing and adaptation to the mechanical micro-environment, has been studied extensively in recent years. A variety of tools for mechanical stimulation have been developed to produce cellular responses. We developed a novel tool, a highly focused ultrasound microbeam, for non-contact cell stimulation at a microscale. This tool, at 200 MHz, was applied to human umbilical vein endothelial cells to investigate its potential to elicit an elevation in cytoplasmic Ca(2+) levels. It was found that the response was dose dependent, and moreover, extracellular Ca(2+) and cytoplasmic Ca(2+) stores were involved in the Ca(2+) elevation. These results suggest that high-frequency ultrasound microbeam stimulation is potentially a novel non-contact tool for studying cellular mechanotransduction if the acoustic pressures at such high frequencies can be quantified.

Systematic study of high-frequency ultrasonic transducer design for laser-scanning photoacoustic ophthalmoscopy.

Photoacoustic ophthalmoscopy (PAOM) is a high-resolution in vivo imaging modality that is capable of providing specific optical absorption information for the retina. A high-frequency ultrasonic transducer is one of the key components in PAOM, which is in contact with the eyelid through coupling gel during imaging. The ultrasonic transducer plays a crucial role in determining the image quality affected by parameters such as spatial resolution, signal-to-noise ratio, and field of view. In this paper, we present the results of a systematic study on a high-frequency ultrasonic transducer design for PAOM. The design includes piezoelectric material selection, frequency selection, and the fabrication process. Transducers of various designs were successfully applied for capturing images of biological samples in vivo. The performances of these designs are compared and evaluated.

Laser-scanning photoacoustic microscopy with ultrasonic phased array transducer.

In this paper, we report our latest progress on proving the concept that ultrasonic phased array can improve the detection sensitivity and field of view (FOV) in laser-scanning photoacoustic microscopy (LS-PAM). A LS-PAM system with a one-dimensional (1D) ultrasonic phased array was built for the experiments. The 1D phased array transducer consists of 64 active elements with an overall active dimension of 3.2 mm × 2 mm. The system was tested on imaging phantom and mouse ear in vivo. Experiments showed a 15 dB increase of the signal-to-noise ratio (SNR) when beamforming was employed compared to the images acquired with each single element. The experimental results demonstrated that ultrasonic phased array can be a better candidate for LS-PAM in high sensitivity applications like ophthalmic imaging.

Early detection of neurodegeneration in brain ischemia by manganese-enhanced MRI.

This study aims to employ in vivo manganese-enhanced MRI (MEMRI) to detect neurodegenerative changes in two models of brain ischemia, photothrombotic cortical injury (PCI) and transient middle cerebral artery occlusion (MCAO) in rodents. After systemic Mn(2+) injection to both ischemic models, a close pattern of T1-weighted hyperintensity was observed throughout different brain regions in comparison to the distribution of GFAP, MnSOD and GS immunoreactivities, whereby conventional MRI could hardly detect such. In addition, the infarct volumes in the posterior parts of the brain had significantly reduced after Mn(2+) injection to the MCAO model. It is suggested that exogenous Mn(2+) injection may provide enhanced MEMRI detection of oxidative stress and gliosis early after brain ischemia. Manganese may also mediate infarctions at remote brain regions in transient focal cerebral ischemia before delayed secondary damage takes place.