Ultrasound Imaging Transducer Research Articles

A Wearable Dual-Mode Probe for Image-Guided Closed-Loop Ultrasound Neuromodulation.

Low-intensity focused ultrasound (FUS) is an emerging non-invasive and spatially/temporally precise method for modulating the firing rates and patterns of peripheral nerves. This paper describes an image-guided platform for chronic and patient-specific FUS neuromodulation. The system uses custom wearable probes containing separate ultrasound imaging and modulation transducer arrays realized using piezoelectric transducers assembled on a flexible printed circuit board (PCB). Dual-mode probes operating around 4 MHz (imaging) and 1.3 MHz (modulation) were fabricated and tested on tissue phantoms. The resulting B-mode images were analyzed using a template-matching algorithm to estimate the location of the target nerve and then direct the modulation beam toward the target. The ultrasound transmit voltage used to excite the modulation array was optimized in real-time by automatically regulating functional feedback signals (the average rates of emulated muscle twitches detected by an on-board motion sensor) through a proportional and integral (PI) controller, thus providing robustness to inter-subject variability and probe positioning errors. The proposed closed-loop neuromodulation paradigm was experimentally demonstrated in vitro using an active tissue phantom that integrates models of the posterior tibial nerve and nearby blood vessels together with embedded sensors and actuators.

A Multiplexed 32 × 32 2D Matrix Array Transducer for Flexible Sub-Aperture Volumetric Ultrasound Imaging.

A fully-sampled two-dimensional (2D) matrix array ultrasonic transducer is essential for fast and accurate three-dimensional (3D) volumetric ultrasound imaging. However, these arrays, usually consisting of thousands of elements, not only face challenges of poor performance and complex wiring due to high-density elements and small element sizes but also put high requirements for electronic systems. Current commercially available fully-sampled matrix arrays, dividing the aperture into four fixed sub-apertures to reduce system channels through multiplexing are widely used. However, the fixed sub-aperture configuration limits imaging flexibility and the gaps between sub-apertures lead to reduced imaging quality. In this study, we propose a high-performance multiplexed matrix array by the design of 1-3 piezocomposite and gapless sub-aperture configuration, as well as optimized matching layer materials. Furthermore, we introduce a sub-aperture volumetric imaging method based on the designed matrix array, enabling high-quality and flexible 3D ultrasound imaging with a low-cost 256-channel system. The influence of imaging parameters, including the number of sub-apertures and steering angle on imaging quality was investigated by simulation, in vitro and in vivo imaging experiments. The fabricated matrix array has a center frequency of 3.4 MHz and a -6 dB bandwidth of above 70%. The proposed sub-aperture volumetric imaging method demonstrated a 10% improvement in spatial resolution, a 19% increase in signal-to-noise ratio, and a 57.7% increase in contrast-to-noise ratio compared with the fixed sub-aperture array imaging method. This study provides a new strategy for high-quality volumetric ultrasound imaging with a low-cost system.

Separate emission/reception transducers for 3D ultrafast ultrasound imaging

Recent advancements in ultrafast 3D ultrasound imaging have revolutionized the field of echography, enabling its extension to novel applications such as cerebral dynamics, cardiac electrophysiology and the quantitative imaging of intrinsic mechanical properties of tumors. To facilitate these advancements, two primary transducer strategies are employed in 3D imaging. The first involves dense 2D probes equipped with a large number of elements, typically exceeding 1024. The second strategy uses row-column addressing, which simplifies the electronic control of the probe elements. Despite their effectiveness, these methods entail complexities in design and fabrication. Addressing these challenges, our study introduces an innovative transducer configuration that distinctly separates emission and reception functionalities. This separation not only simplifies the overall transducer design but also significantly reduces the system's complexity. A sparse array of PVDF transducers, which have been laser micro-machined to ensure acoustic transparency, is used at the reception. For the emission aspect, we employ a specialized acoustic concentrator to emulate point-like emission. The paper presents the detailed design requirements, assembly process, and the operational principles of this novel transducer. Furthermore, an experimental validation is conducted using a CIRS 040GSE phantom model. This validation is crucial to demonstrate the practical applicability and reliability of our transducer in real-world medical imaging scenarios.

An ultrasensitive and broadband transparent ultrasound transducer for ultrasound and photoacoustic imaging in-vivo

Transparent ultrasound transducers (TUTs) can seamlessly integrate optical and ultrasound components, but acoustic impedance mismatch prohibits existing TUTs from being practical substitutes for conventional opaque ultrasound transducers. Here, we propose a transparent adhesive based on a silicon dioxide-epoxy composite to fabricate matching and backing layers with acoustic impedances of 7.5 and 4–6 MRayl, respectively. By employing these layers, we develop an ultrasensitive, broadband TUT with 63% bandwidth at a single resonance frequency and high optical transparency ( > 80%), comparable to conventional opaque ultrasound transducers. Our TUT maximises both acoustic power and transfer efficiency with maximal spectrum flatness while minimising ringdowns. This enables high contrast and high-definition dual-modal ultrasound and photoacoustic imaging in live animals and humans. Both modalities reach an imaging depth of > 15 mm, with depth-to-resolution ratios exceeding 500 and 370, respectively. This development sets a new standard for TUTs, advancing the possibilities of sensor fusion.

Towards a locally constructed multi-element ultrasound imaging transducer for resource poor environments

This paper investigates techniques and materials for making a multi-element ultrasound imaging transducer with craft-based techniques available in resource poor environments. The transducer housing can be conveniently divided into three parts: the body supporting the piezoelectric (PZT) elements and other components; the matching layer between the PZT elements and the human body; and the backing layer behind the PZT elements. Low-cost 3D printing systems based on photopolymers were found to be suitable for manufacturing the body. Finite Element Modelling (FEM) showed that the material characteristics of the backing layer and the thickness of the matching layer were much less critical than predicted by ultrasound plane wave theory and transmission line theory, respectively. The backing and matching layers are normally made from epoxy-tungsten composites that are pourable in the uncured state. However, the composite required for the backing layer was putty-like when uncured. When the tungsten was allowed to settle under gravity during curing, a 20 % by volume uncured tungsten-epoxy composite gave a 30 % by volume concentration of tungsten at the bottom when cured at 20–30 °C. These findings, when coupled with the findings from the FEM modelling, suggests that constructing a multi-element ultrasound imaging transducer using craft-based techniques is feasible.

Curved toroidal row column addressed transducer for 3D ultrafast ultrasound imaging.

3D Imaging of the human heart at high frame rate is of major interest for various clinical applications. Electronic complexity and cost has prevented the dissemination of 3D ultrafast imaging into the clinic. Row column addressed (RCA) transducers provide volumetric imaging at ultrafast frame rate by using a low electronic channel count, but current models are ill-suited for transthoracic cardiac imaging due to field-of-view limitations. In this study, we proposed a mechanically curved RCA with an aperture adapted for transthoracic cardiac imaging (24 × 16 mm²). The RCA has a toroidal curved surface of 96 elements along columns (curvature radius rC = 4.47 cm) and 64 elements along rows (curvature radius rR = 3 cm). We implemented delay and sum beamforming with an analytical calculation of the propagation of a toroidal wave which was validated using simulations (Field II). The imaging performance was evaluated on a calibrated phantom. Experimental 3D imaging was achieved up to 12 cm deep with a total angular aperture of 30° for both lateral dimensions. The Contrast-to-Noise ratio increased by 12 dB from 2 to 128 virtual sources. Then, 3D Ultrasound Localization Microscopy (ULM) was characterized in a sub-wavelength tube diameter. Finally, 3D ULM was demonstrated on a perfused ex-vivo swine heart to image the coronary microcirculation.

An electrical impedance matching method of dual-frequency transducers for ultrasound internal imaging

A micro single transducer with two resonant frequencies can solve the inherent contradiction between detection depth and resolution by simultaneously emitting two frequency bands of ultrasonic waves. But the difference of impedance values in the two resonant frequency bands seriously affects dual-frequency echo signal efficiency. In order to improve the quality of transducer sensitivity, a dual-frequency electrical impedance matching method is proposed in the study. By selecting a specific matching structure to match one frequency without messing up the impedance of another frequency, the two frequencies can be designed independently to achieve preliminary impedance matching. The matching networks for single frequency are combined to form a complete dual-frequency matching circuit, and adjust the initial matching value to obtain a suitable matching state. The performances of the matching network have been tested and verified by experiments. After matching, the low-frequency and high-frequency sensitivities of the transducer increased 1.86 times and 2.26 times, respectively. In addition, system noise was also reduced by 29.6% after adding the matching network. The results demonstrate that this dual-frequency matching network can simultaneously increase the amplitude of high-frequency and low-frequency echoes to obtain better quality images.

A Scholte wave approach for ultrasonic surface acoustic wave elastography.

Pathological changes in tissues are often related to changes in tissue mechanical properties, making elastography an important tool for medical applications. Among the existing elastography methods, ultrasound elastography is of great interest due to the inherent advantages of ultrasound imaging technology, such as low cost, portability, safety, and wide availability. Although ultrasonic shear wave elastography, as a platform technology, can potentially quantify the elasticity of tissue at any depth, its current implementation cannot assess superficial tissue but can only image deep tissue. To address this challenge, we proposed an ultrasonic Scholte-wave-based approach for imaging the elasticity of superficial tissue. The feasibility of the proposed technique was tested using a gelatin phantom with a cylindrical inclusion. To generate Scholte wave in the superficial region of the phantom, we proposed a new experimental configuration in which a liquid layer was introduced between an ultrasound imaging transducer and the tissue-mimicking phantom. We utilized an acoustic radiation force impulse to excite the tissue-mimicking phantom, analyzed the properties of the generated Scholte waves, and applied the waves for elasticity imaging. In this study, we first reported the observation that Scholte (surface) waves and shear (bulk) waves were simultaneously generated, and they propagated in the superficial and deeper regions of the phantom, respectively. Then, we presented some important properties of the generated Scholte waves. For a 5w/v% gelatin phantom, the generated Scholte waves have a speed of around 0.9m/s, a frequency of about 186Hz, and thus a wavelength of about 4.8mm. The speed ratio between the simultaneously generated Scholte wave and shear wave is about 0.717, which is 15% lower than the theoretical expectation. And we further demonstrated the feasibility of Scholte wave as a mechanism for imaging superficial tissue elasticity. Together with the simultaneously generated shear wave, the Scholte wave was shown to be able to quantitatively image both the background and the cylindrical inclusion (4mm in diameter) of the tissue-mimicking gelatin phantom. This work shows that the elasticity of superficial tissue can be evaluated by utilizing the generated Scholte wave alone, and it also shows that a comprehensive elasticity imaging of the tissue extending from the superficial to deep regions can be achieved by combining the proposed Scholte wave technique and the conventional shear wave technique.

Sonodynamic Therapy With Concentric Ultrasound Imaging Array for Precision Theranostics for Atherosclerotic Plaque.

Atherosclerotic cardiovascular disease is a major cause of human disability and mortality. Our previous study demonstrated the safety and efficacy of sonodynamic therapy (SDT) on atherosclerotic plaques. However, traditional single-element therapeutic transducer has single acoustic field, and positioning therapeutic and imaging transducers in the same position is difficult during ultrasound imaging-guided SDT. Continuously changing the position of transducers to intervene lesions in different positions is required, increasing the difficulty of treatment. Thus, an SDT device with precise theranostics is required. Therefore, we designed and fabricated a "concentric ultrasound transducer for theranostics" (CUST-T), comprising a central 8-MHz linear array transducer for ultrasound imaging, and a peripheral 1-MHz hollow two-dimensional (2-D) planar array transducer for generating phased-array focused ultrasound (PAFUS). The CUST-T exhibited high imaging resolution at a distance of up to 20 mm from the transducer and could generate a personalized complex PAFUS acoustic field to match various lesions. In vitro biomedical results showed that PAFUS-SDT induced RAW264.7-derived foam cell apoptosis leading to a targeting field apoptotic rate 4.36-6.24 times that of the nontargeting field and the significant apoptotic region was consistent with the PAFUS acoustic field. In vivo, PAFUS-SDT guided by ultrasound imaging significantly increased the lumen area ( ) and collagen level ( ), whereas the wall thickness ( ) and lipid content ( ) of rabbit femoral artery were reduced. In conclusion, CUST-T provided image guidance sufficient for accurate SDT for atherosclerotic plaques in peripheral arteries and could be applied in clinical practice.

Palpating particles using the acoustic radiation force: A new approach to magnetic particle imaging

Magnetic particle imaging (MPI) is a tracer-based imaging technique that does not require radiation or a cyclotron. However, it does require bulky MRI-like hardware and does not produce a background tissue image. We propose a new approach to MPI that uses the acoustic radiation force to dynamically move iron nanoparticles through a magnetic field gradient, thereby changing their magnetic moment and producing a detectable signal in a receiver coil. An image could be formed by electronically steering the ultrasound focus to push particles at different spatial locations, like an ultrasonic palpation. This would enable a device that attaches to an ultrasound imaging transducer for simultaneous MPI and inherently registered ultrasound imaging. To test the feasibility of the technique, a cylindrical agar and graphite phantom was placed between a Maxwell coil pair which generated a linear field gradient across it, and particle signals were detected using a receiver solenoid. 2-ms ultrasound pulses were applied using a 6 MHz transducer to generate displacements along the magnetic field gradient. Signal measurements confirmed a lack of signal when iron nanoparticles were absent from the phantom, but the signal was detected when particles were present, with amplitudes that linearly increased with the gradient coil current.

Potassium Sodium Niobate-Based Lead-Free High-Frequency Ultrasonic Transducers for Multifunctional Acoustic Tweezers

Ultrasonic transducers may need to operate in direct contact with the human body, especially with the skin or closer to blood vessels. Eco-friendly lead-free materials and devices are therefore being vigorously developed for biosafety considerations. This work presents high-performance potassium sodium niobate [(K,Na)NbO3, KNN]-based lead-free ceramics with composition-driven multiphase coexistence and their application on high-frequency ultrasonic transducers for multifunctional acoustic tweezers. A high piezoelectric constant d33 value of 332 pC/N, a good Curie temperature TC value of 348 °C, and improved in situ temperature stability were obtained in the piezoceramics via the construction multiple phases near room temperature and domain engineering. One to three piezocomposites were further fabricated based on the synthesized ceramics for higher electromechanical coupling properties. Lead-free high-frequency transducers as multifunctional acoustic tweezers for precise and selective manipulation of microparticles were designed and manufactured with a high center frequency of 23.4 MHz and a broad -6 dB bandwidth of 75.4%. Additionally, a stable transducer performance was obtained over a test temperature range of 23-60 °C, indicating good thermal stability in environments with fluctuating temperatures. Research on lead-free high-frequency transducers for ultrasound imaging and precise and selective manipulation of microparticles demonstrates their broad potential in fields such as medical therapy and diagnosis.

High-Attenuation Backing Layer for Miniaturized Ultrasound Imaging Transducer.

Current miniaturized ultrasound transducers suffer from insufficient attenuation from the backing layer due to their limited thickness. The thickness of the backing layer is one of the critical factors determining the device size and transducer performance for miniaturized transducers inserted and operated in a limited space. Glass bubbles, polyamide resin, and tungsten powder are combined to form a new highly attenuative backing material. It has high attenuation (>160 dB/cm at 5 MHz), which is five times greater than silver-based conductive epoxy commonly used for high-frequency ultrasound transducers, appropriate acoustic impedance (4.6 MRayl), and acceptable damping capability. An intravascular ultrasound (IVUS) transducer constructed with the 170 [Formula: see text] of the proposed backing layer demonstrated that the amplitude of the signal returned from the backing layer was 1.8 times smaller, with ring-down attenuated by 6 dB. Wire-phantom imaging revealed that the axial resolution was 30% better with the suggested backing than silver-based conductive epoxy backing. Because of its excellent attenuation capability even at a limited thickness, simple manufacturing process, and easy customization capability, the suggested highly attenuative backing layer may be used for miniaturized ultrasound transducers.

Fiber Optic Photoacoustic-Based Steering Imaging Transducer

This letter presents a novel fiber optic imaging transducer for solving ultrasound steering problems for photoacoustic imaging (PAI). The design of the transducer is explored and verified by photoacoustic (PA) effect and Fabry-Perot (FP) principle. A directional ultrasound beam that is based on PA effect is excited by coating gold nanocomposites with polydimethylsiloxane (PDMS) at the distal end of an optical fiber with a certain degree of offset. A microlens with a concave surface is attached to the surface to focus on the length of ultrasound waves. A pulse echo signal is collected by an FP fiber optic sensor. Ultrasound field distribution is captured in order to confirm that the ultrasound beam is directional and focused as a result of the microlens. B mode imaging tests on <italic xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">ex vivo</i> swine back rib muscle-fat slices report the transducer to own a higher depth of penetration. Ultrasound imaging is formed by combining the scanned pulse echo signal without the need for an image reconstruction algorithm. The results demonstrate a promising foundation for the prototype of the fiber optic ultrasound imaging transducer in terms of challenging environments and in <italic xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">vivo</i> .

Dual-Use Transducer for Ultrasound Imaging and Pulsed Focused Ultrasound Therapy.

Pulsed focused ultrasound (pFUS) uses short acoustic pulses delivered at low duty cycle and moderate intensity to noninvasively apply mechanical stress or introduce disruption to tissue. Ultrasound-guided pFUS has primarily been used for inducing cavitation at the focus, with or without contrast agents, to promote drug delivery to tumors. When applied in tandem with contrast agents, pFUS is often administered using an ultrasound imaging probe, which has a small footprint and does not require a large acoustic window. The use of nonlinear pFUS without contrast agents was recently shown to be beneficial for localized tissue disruption, but required higher ultrasound pressure levels than a conventional ultrasound imaging probe could produce. In this work, we present the design of a compact dual-use 1-MHz transducer for ultrasound-guided pFUS without contrast agents. Nonlinear pressure fields that could be generated by the probe, under realistic power input, were simulated using the Westervelt equation. In water, fully developed shocks of 42-MPa amplitude and peak negative pressure of 8 MPa were predicted to form at the focus at 458-W acoustic power or 35% of the maximum reachable power of the transducer. In absorptive soft tissue, fully developed shocks formed at higher power (760 W or 58% of the maximum reachable power) with the shock amplitude of 33 MPa and peak negative pressure of 7.5 MPa. The electronic focus-steering capabilities of the array were evaluated and found to be sufficient to cover a target with dimensions of 19 mm in axial direction and 44 mm in transversal direction.

Homogenized electromechanical coefficients and effective parameters of 1–3 piezocomposites for ultrasound imaging transducers

Analytical models can be useful tools to develop efficient transducers dedicated to a specific application field. This work proposes a homogenization technique for establishing the effective coefficients and parameters of 1-3 piezocomposite whose both phases are piezoelectrically active. Such piezocomposite materials may have homogenized electromechanical properties resulting from positive hybrid effect. A numerical simulation of the effective parameters resulting from a PZT-5A / PVDF-TrFE composition shows that, volume fraction influences significantly on the properties of the piezocomposite and consequently on its behaviour in service. As an end-user application for ultrasound imaging, those piezocomposite materials are effectively integrated in a piezocomposite transducer, coupled with a dedicated backing. The resulting characteristics in terms of impulse response and associated electroacoustic echo are compared and discussed for typical configurations. A method for the optimization of the design of such piezocomposite transducer is presented. Specific estimators based on the bandwidth flatness BWF and bandwidth amplitude product BWA are proposed as stable and smooth criteria for an optimization procedure.

Lead halide perovskite for efficient optoacoustic conversion and application toward high-resolution ultrasound imaging

Lead halide perovskites have exhibited excellent performance in solar cells, LEDs and detectors. Thermal properties of perovskites, such as heat capacity and thermal conductivity, have rarely been studied and corresponding devices have barely been explored. Considering the high absorption coefficient (104~105 cm−1), low specific heat capacity (296–326 J kg−1 K−1) and small thermal diffusion coefficient (0.145 mm2 s−1), herein we showcase the successful use of perovskite in optoacoustic transducers. The theoretically calculated phonon spectrum shows that the overlap of optical phonons and acoustic phonons leads to the up-conversion of acoustic phonons, and thus results in experimentally measured low thermal diffusion coefficient. The assembled device of PDMS/MAPbI3/PDMS simultaneously achieves broad bandwidths (−6 dB bandwidth: 40.8 MHz; central frequency: 29.2 MHz), and high conversion efficiency (2.97 × 10−2), while all these parameters are the record values for optoacoustic transducers. We also fabricate miniatured devices by assembling perovskite film onto fibers, and clearly resolve the fine structure of fisheyes, which demonstrates the strong competitiveness of perovskite based optoacoustic transducers for ultrasound imaging.

Recent Advances in Transducers for Intravascular Ultrasound (IVUS) Imaging.

As a well-known medical imaging methodology, intravascular ultrasound (IVUS) imaging plays a critical role in diagnosis, treatment guidance and post-treatment assessment of coronary artery diseases. By cannulating a miniature ultrasound transducer mounted catheter into an artery, the vessel lumen opening, vessel wall morphology and other associated blood and vessel properties can be precisely assessed in IVUS imaging. Ultrasound transducer, as the key component of an IVUS system, is critical in determining the IVUS imaging performance. In recent years, a wide range of achievements in ultrasound transducers have been reported for IVUS imaging applications. Herein, a comprehensive review is given on recent advances in ultrasound transducers for IVUS imaging. Firstly, a fundamental understanding of IVUS imaging principle, evaluation parameters and IVUS catheter are summarized. Secondly, three different types of ultrasound transducers (piezoelectric ultrasound transducer, piezoelectric micromachined ultrasound transducer and capacitive micromachined ultrasound transducer) for IVUS imaging are presented. Particularly, the recent advances in piezoelectric ultrasound transducer for IVUS imaging are extensively examined according to their different working mechanisms, configurations and materials adopted. Thirdly, IVUS-based multimodality intravascular imaging of atherosclerotic plaque is discussed. Finally, summary and perspectives on the future studies are highlighted for IVUS imaging applications.

Thermal Ablation and High-Resolution Imaging Using a Back-to-Back (BTB) Dual-Mode Ultrasonic Transducer: In Vivo Results.

We present a back-to-back (BTB) structured, dual-mode ultrasonic device that incorporates a single-element 5.3 MHz transducer for high-intensity focused ultrasound (HIFU) treatment and a single-element 20.0 MHz transducer for high-resolution ultrasound imaging. Ultrasound image-guided surgical systems have been developed for lesion monitoring to ensure that ultrasonic treatment is correctly administered at the right locations. In this study, we developed a dual-element transducer composed of two elements that share the same housing but work independently with a BTB structure, enabling a mode change between therapy and imaging via 180-degree mechanical rotation. The optic fibers were embedded in the HIFU focal region of ex vivo chicken breasts and the temperature change was measured. Images were obtained in vivo mice before and after treatment and compared to identify the treated region. We successfully acquired B-mode and C-scan images that display the hyperechoic region indicating coagulation necrosis in the HIFU-treated volume up to a depth of 10 mm. The compact BTB dual-mode ultrasonic transducer may be used for subcutaneous thermal ablation and monitoring, minimally invasive surgery, and other clinical applications, all with ultrasound only.

The Preparation of Chicken Ex Ovo Embryos and Chorioallantoic Membrane Vessels as In Vivo Model for Contrast-Enhanced Ultrasound Imaging and Microbubble-Mediated Drug Delivery Studies.

The chicken embryo and the blood-vessel rich chorioallantoic membrane (CAM) is a valuable in vivo model to investigate biomedical processes, new ultrasound pulsing schemes, or novel transducers for contrast-enhanced ultrasound imaging and microbubble-mediated drug delivery. The reasons for this are the accessibility of the embryo and vessel network of the CAM as well as the low costs of the model. An important step to get access to the embryo and CAM vessels is to take the egg content out of the eggshell. In this protocol, three methods for taking the content out of the eggshell between day 5 and8 of incubation are described thus allowing the embryos to develop inside the eggshell up to these days. The described methods only require simple tools and equipment and yield a higher survival success rate of 90% for 5-day, 75% for 6-day, 50% for 7-day, and 60% for 8-day old incubated eggs in comparison to ex ovo cultured embryos (~50%). The protocol also describes how to inject cavitation nuclei, such as microbubbles, into the CAM vascular system, how to separate the membrane containing the embryo and CAM from the rest of the egg content for optically transparent studies, and how to use the chicken embryo and CAM in a variety of short-term ultrasound experiments. The in vivochicken embryo and CAM model is extremely relevant to investigate novel imaging protocols, ultrasound contrast agents, and ultrasound pulsing schemes for contrast-enhanced ultrasound imaging, and to unravel the mechanisms of ultrasound-mediated drug delivery.

The Preparation of Chicken Ex Ovo Embryos and Chorioallantoic Membrane Vessels as In Vivo Model for Contrast-Enhanced Ultrasound Imaging and Microbubble-Mediated Drug Delivery Studies

The chicken embryo and the blood-vessel rich chorioallantoic membrane (CAM) is a valuable in vivo model to investigate biomedical processes, new ultrasound pulsing schemes, or novel transducers for contrast-enhanced ultrasound imaging and microbubble-mediated drug delivery. The reasons for this are the accessibility of the embryo and vessel network of the CAM as well as the low costs of the model. An important step to get access to the embryo and CAM vessels is to take the egg content out of the eggshell. In this protocol, three methods for taking the content out of the eggshell between day 5 and 8 of incubation are described thus allowing the embryos to develop inside the eggshell up to these days. The described methods only require simple tools and equipment and yield a higher survival success rate of 90% for 5-day, 75% for 6-day, 50% for 7-day, and 60% for 8-day old incubated eggs in comparison to ex ovo cultured embryos (~50%). The protocol also describes how to inject cavitation nuclei, such as microbubbles, into the CAM vascular system, how to separate the membrane containing the embryo and CAM from the rest of the egg content for optically transparent studies, and how to use the chicken embryo and CAM in a variety of short-term ultrasound experiments. The in vivo chicken embryo and CAM model is extremely relevant to investigate novel imaging protocols, ultrasound contrast agents, and ultrasound pulsing schemes for contrast-enhanced ultrasound imaging, and to unravel the mechanisms of ultrasound-mediated drug delivery.