Search

The potential of proton therapy to improve the conformity of the delivered dose to the tumor volume is currently limited by range uncertainties. Injectable superheated nanodroplets have recently been proposed for ultrasound-based in vivo range verification, as these vaporize into echogenic microbubbles on proton irradiation. In previous studies, offline ultrasound images of phantoms with dispersed nanodroplets were acquired after irradiation, relating the induced vaporization profiles to the proton range. However, the aforementioned method did not enable the counting of individual vaporization events, and offline imaging cannot provide real-time feedback. In this study, we overcame these limitations using high-frame-rate ultrasound imaging with a linear array during proton irradiation of phantoms with dispersed perfluorobutane nanodroplets at 37°C and 50°C. Differential image analysis of subsequent frames allowed us to count individual vaporization events and to localize them with a resolution beyond the ultrasound diffraction limit, enabling spatial and temporal quantification of the interaction between ionizing radiation and nanodroplets. Vaporization maps were found to accurately correlate with the stopping distribution of protons (at 50°C) or secondary particles (at both temperatures). Furthermore, a linear relationship between the vaporization count and the number of incoming protons was observed. These results indicate the potential of real-time high-frame-rate contrast-enhanced ultrasound imaging for proton range verification and dosimetry., Competing Interests: Conflict of interest disclosure The authors declare no conflict of interest., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Volumetric ultrasound imaging of blood flow with microbubbles enables a more complete visualization of the microvasculature. Sparse arrays are ideal candidates to perform volumetric imaging at reduced manufacturing complexity and cable count. However, due to the small number of transducer elements, sparse arrays often come with high clutter levels, especially when wide beams are transmitted to increase the frame rate. In this study, we demonstrate with a prototype sparse array probe and a diverging wave transmission strategy, that a uniform transmission field can be achieved. With the implementation of a spatial coherence beamformer, the background clutter signal can be effectively suppressed, leading to a signal to background ratio improvement of 25 dB. With this approach, we demonstrate the volumetric visualization of single microbubbles in a tissue-mimicking phantom as well as vasculature mapping in a live chicken embryo chorioallantoic membrane.

High-frame-rate (HFR) echo-particle image velocimetry (echoPIV) is a promising tool for measuring intracardiac blood flow dynamics. In this study, we investigate the optimal ultrasound contrast agent (UCA: SonoVue) infusion rate and acoustic output to use for HFR echoPIV (PRF = 4900 Hz) in the left ventricle (LV) of patients. Three infusion rates (0.3, 0.6, and 1.2 ml/min) and five acoustic output amplitudes (by varying transmit voltage: 5, 10, 15, 20, and 30 V-corresponding to mechanical indices of 0.01, 0.02, 0.03, 0.04, and 0.06 at 60-mm depth) were tested in 20 patients admitted for symptoms of heart failure. We assess the accuracy of HFR echoPIV against pulsed-wave Doppler acquisitions obtained for mitral inflow and aortic outflow. In terms of image quality, the 1.2-ml/min infusion rate provided the highest contrast-to-background ratio (CBR) (3-dB improvement over 0.3 ml/min). The highest acoustic output tested resulted in the lowest CBR. Increased acoustic output also resulted in increased microbubble disruption. For the echoPIV results, the 1.2-ml/min infusion rate provided the best vector quality and accuracy; mid-range acoustic outputs (corresponding to 15-20-V transmit voltages) provided the best agreement with the pulsed-wave Doppler. Overall, the highest infusion rate (1.2 ml/min) and mid-range acoustic output amplitudes provided the best image quality and echoPIV results.

Microbubble translations driven by ultrasound-induced radiation forces can be beneficial for applications in ultrasound molecular imaging and drug delivery. Here, the effect of size range in microbubble populations on their translations is investigated experimentally and theoretically. The displacements within five distinct size-isolated microbubble populations are driven by a standard ultrasound-imaging probe at frequencies ranging from 3 to 7 MHz, and measured using the multi-gate spectral Doppler approach. Peak microbubble displacements, reaching up to 10 μm per pulse, are found to describe transient phenomena from the resonant proportion of each bubble population. The overall trend of the statistical behavior of the bubble displacements, quantified by the total number of identified displacements, reveals significant differences between the bubble populations as a function of the transmission frequency. A good agreement is found between the experiments and theory that includes a model parameter fit, which is further supported by separate measurements of individual microbubbles to characterize the viscoelasticity of their stabilizing lipid shell. These findings may help to tune the microbubble size distribution and ultrasound transmission parameters to optimize the radiation-force translations. They also demonstrate a simple technique to characterize the microbubble shell viscosity, the fitted model parameter, from freely floating microbubble populations using a standard ultrasound-imaging probe.

Due to the primary radiation force, microbubble displacement has been observed previously in the focal region of single-element and array ultrasound probes. This effect has been harnessed to increase the contact between the microbubbles and targeted endothelium for drug delivery and ultrasound molecular imaging. In this study, microbubble displacements associated with plane-wave (PW) transmission are thoroughly investigated and compared to those obtained in focused-wave (FW) transmission over a range of pulse repetition frequencies, burst lengths (BLs), peak negative pressures, and transmission frequencies. In PW mode, the displacements, depending upon the experimental conditions, are in some cases consistently higher (e.g., by 28%, when the longest BL was used at PRF = 4 kHz), and the axial displacements are spatially more uniform compared to FW mode. Statistical analysis on the measured displacements reveals a slightly different frequency dependence of statistical quantities compared to transient peak microbubble displacements, which may suggest the need to consider the size range within the tested microbubble population.

Acoustic behavior of lipid-coated microbubbles has been widely studied, which has led to several numerical microbubble dynamics models that incorporate lipid coating behavior, such as buckling and rupture. In this study we investigated the relationship between microbubble acoustic and lipid coating behavior on a nanosecond scale by using fluorescently labeled lipids. It is hypothesized that a local increased concentration of lipids, appearing as a focal area of increased fluorescence intensity (hot spot) in the fluorescence image, is related to buckling and folding of the lipid layer thereby highly influencing the microbubble acoustic behavior. To test this hypothesis, the lipid microbubble coating was fluorescently labeled. The vibration of the microbubble (n = 177; 2.3-10.3 μm in diameter) upon insonification at an ultrasound frequency of 0.5 or 1 MHz at 25 or 50 kPa acoustic pressure was recorded with the UPMC Cam, an ultra-high-speed fluorescence camera, operated at ~4-5 million frames per second. During short tone-burst excitation, hot spots on the microbubble coating occurred at relative vibration amplitudes > 0.3 irrespective of frequency and acoustic pressure. Around resonance, the majority of the microbubbles formed hot spots. When the microbubble also deflated acoustically, hot spot formation was likely irreversible. Although compression-only behavior (defined as substantially more microbubble compression than expansion) and subharmonic responses were observed in those microbubbles that formed hot spots, both phenomena were also found in microbubbles that did not form hot spots during insonification. In conclusion, this study reveals hot spot formation of the lipid monolayer in the microbubble's compression phase. However, our experimental results show that there is no direct relationship between hot spot formation of the lipid coating and microbubble acoustic behaviors such as compression-only and the generation of a subharmonic response. Hence, our hypothesis that hot spots are related to acoustic buckling could not be verified.

Lipid-coated microbubbles are used clinically as contrast agents for ultrasound imaging and are being developed for a variety of therapeutic applications. The lipid encapsulation and shedding of the lipids by acoustic driving of the microbubble has a crucial role in microbubble stability and in ultrasound-triggered drug delivery; however, little is known about the dynamics of lipid shedding under ultrasound excitation. Here we describe a study that optically characterized the lipid shedding behavior of individual microbubbles on a time scale of nanoseconds to microseconds. A single ultrasound burst of 20 to 1000 cycles, with a frequency of 1 MHz and an acoustic pressure varying from 50 to 425 kPa, was applied. In the first step, high-speed fluorescence imaging was performed at 150,000 frames per second to capture the instantaneous dynamics of lipid shedding. Lipid detachment was observed within the first few cycles of ultrasound. Subsequently, the detached lipids were transported by the surrounding flow field, either parallel to the focal plane (in-plane shedding) or in a trajectory perpendicular to the focal plane (out-of-plane shedding). In the second step, the onset of lipid shedding was studied as a function of the acoustic driving parameters, for example, pressure, number of cycles, bubble size and oscillation amplitude. The latter was recorded with an ultrafast framing camera running at 10 million frames per second. A threshold for lipid shedding under ultrasound excitation was found for a relative bubble oscillation amplitude >30%. Lipid shedding was found to be reproducible, indicating that the shedding event can be controlled., (Copyright © 2014 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

Ultrasound contrast agents are valuable in diagnostic ultrasound imaging, and they increasingly show potential for drug delivery. This review focuses on the acoustic behavior of flexible-coated microbubbles and rigid-coated microcapsules and their contribution to enhanced drug delivery. Phenomena relevant to drug delivery, such as non-spherical oscillations, shear stress, microstreaming, and jetting will be reviewed from both a theoretical and experimental perspective. Further, the two systems for drug delivery, co-administration and the microbubble as drug carrier system, are reviewed in relation to the microbubble behavior. Finally, future prospects are discussed that need to be addressed for ultrasound contrast agents to move from a pre-clinical tool into a clinical setting., (Copyright © 2014 Elsevier B.V. All rights reserved.)

A detailed understanding of the response of single microbubbles subjected to ultrasound is fundamental to a full understanding of the contrast-enhancing abilities of microbubbles in medical ultrasound imaging, in targeted molecular imaging with ultrasound, and in ultrasound-mediated drug delivery with microbubbles. Here, single microbubbles are isolated and their ultrasound-induced radial dynamics recorded with an ultra-high-speed camera at up to 25 million frames per second. The sound emission is recorded simultaneously with a calibrated single element transducer. It is shown that the sound emission can be predicted directly from the optically recorded radial dynamics, and vice versa, that the nanometer-scale radial dynamics can be predicted from the acoustic response recorded in the far field.

In this experimental study, the nonspherical and translational behavior of individual coated microbubbles of different sizes, in contact with a 20-μm thickness cellulose wall, are observed and categorized systematically. Images from two orthogonally positioned microscopes are merged and then recorded with an ultra-fast framing camera. Large nonspherical deformations were found with 2.25 MHz frequency ultrasound pulses having driving pressures from 80 to 325 kPa. A parametric model combining potential flow theory with a viscous boundary layer at the wall is developed and used to calculate stresses from the optically recorded microbubble oscillations. Peak shear stress of up to 300 kPa and normal stresses of up to 1 MPa are estimated when microbubbles are insonifed with a 2.25 MHz pulse at 325 kPa. The clinical relevance of these results is discussed., (Copyright © 2011 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

This study investigates the use of an acoustic parametric array as a means for microbubble excitation. The excitation wave is generated during propagation in a nonlinear medium of two high-frequency carrier waves, whereby the frequency of the excitation wave is the difference frequency of the carrier waves. Carrier waves of around 10 and 25 MHz are used to generate low-frequency waves between 0.5 and 3.5 MHz at amplitudes in the range of 25 to 80 kPa in water. We demonstrate with high-speed camera observations that it is possible to induce microbubble oscillations with the low frequency signal arising from the nonlinear propagation process. As an application, we determined the resonance frequency of Definity contrast agent microbubbles with radius ranging from 1.5 to 5 μm by sweeping the difference frequency in the range from 0.5 to 3.5 MHz.

Ultrasound contrast agents (UCA) populations are typically polydisperse and contain microbubbles with radii over a given range. Although the behavior of microbubbles of certain sizes might be masked by the behavior of others, the acoustic characterization of UCA is typically made on full populations. In this paper, we have combined acoustic and optical methods to investigate the response of isolated lipid-shelled microbubbles to low-pressure (49 and 62 kPa peak negative pressure) ultrasound tone bursts. These bursts induced slow deflation of the microbubbles. The experimental setup included a microscope connected to a fast camera acquiring one frame per pulse transmitted by a single-element transducer. The behavior of each bubble was measured at multiple frequencies, by cyclically changing the transmission frequency over the range of 2 to 4 MHz during subsequent pulse repetition intervals. The bubble echoes were captured by a second transducer and coherently recorded. More than 50 individual microbubbles were observed. Microbubbles with radii larger than 3 mum did not experience any size reduction. Smaller bubbles slowly deflated, generally until the radius reached a value around 1.4 microm, independent of the initial microbubble size. The detected pressure amplitude backscattered at 2.5 cm distance was very low, decreasing from about 5 Pa down to 1 Pa at 2 MHz as the bubbles deflated. The resonant radius was evaluated from the echo amplitude normalized with respect to the geometrical cross section. At 2-MHz excitation, deflating microbubbles showed highest normalized echo when the radius was 2.2 microm while at higher excitation frequencies, the resonant radius was lower. The relative phase shift of the echo during the deflation process was also measured. It was found to exceed pi/2 in all cases. A heuristic procedure based on the analysis of multiple bubbles of a same population was used to estimate the undamped natural frequency. It was found that a microbubble of 1.7 microm has an undamped natural frequency of 2 MHz. The difference between this size and the resonant radius is discussed as indicative of significant damping.

Radial modulation imaging is a new promising technique to improve contrast-enhanced ultrasound images. The method is based on dual-frequency insonation of contrast agent microbubbles. A low-frequency (LF) pulse is used to modulate the responses of the microbubbles to a high-frequency (HF) imaging pulse. Inverting the LF pulse induces amplitude and phase differences in the HF response of contrast agent microbubbles, which can be detected using Doppler techniques. Although the technique has been successfully implemented, no consensus persists on parameter choice and resulting effects. In a separate study, "compression-only" behavior of coated microbubbles was observed. Compression-only behavior could be beneficial for radial modulation imaging. This was investigated using high-speed camera recordings and simulations. We recorded the vibrations of 78 single microbubbles in a dual-frequency ultrasound field. The results showed that the LF pulse induced significant compression-only behavior, which for microbubble sizes below and at HF resonance resulted in high radial amplitude modulation. It, however, also appeared that, for radial modulation imaging, microbubble size is more important than resonance and compression-only effects.

Ultrasound contrast agents (UCAs) are widely used in Doppler studies, either for simple echo enhancement purposes, or to increase the low signal-to-clutter ratio typical of microcirculation investigations. Common to all Doppler techniques, which are briefly reviewed in this paper, is the basic assumption that possible phase and amplitude changes in received echoes are only associated with UCA microbubble movements due to the drag force of blood. Actually, when UCAs are insonified, phenomena such as rupture, displacement due to radiation force, and acoustically driven deflation might influence the results of Doppler investigations. In this paper, we investigate the possible Doppler effects of such phenomena by means of a numerical simulation model and a special acousto-optical set-up which allows analysis of the behavior of individual microbubbles over relatively long time intervals. It is thus found that all phenomena produce evident Doppler effects in vitro, but that bubble displacement and deflation in particular, are not expected to significantly interfere with clinical measurements in standard conditions.

Radially oscillating microbubbles can deform when in contact with a wall. These nonspherical shapes have a preferential orientation perpendicular to the wall. Conventional microscope setups for microbubble studies have their optical axis perpendicular to the wall (top view); consequently they have a limited view of the deformation of the bubble. We developed a method to image the bubble in a side view by integrating a mirror in the microscope setup. The image was recorded at 14.5 million frames per second by a high-speed camera. When insonified by a 1-MHz, 140-kPa ultrasound pulse, a 9-microm diameter coated bubble appeared spherical in the top view, but strongly nonspherical in the side view. Its shape was alternatively oblate and prolate, with maximum second order spherical harmonic amplitude equal to the radius.

Recent optical and acoustical studies have shown a threshold behaviour in the response of phospholipid-coated contrast agents, for a certain range of sizes. Below the acoustic pressure threshold, the microbubbles' scattering efficacy is significantly reduced compared to that above the threshold. Here we investigate the clinical relevance of the observed threshold behaviour. A cardiac ultrasound scanner system was used to analyse the pressure-dependence of the scatter intensity. The scattering of a native suspension of a phospholipid-coated contrast agent was compared to that of a suspension in which microbubbles with a size larger than 3.0 microm in diameter were extracted. A power modulation scheme at the fundamental frequency was applied. After linearly scaling and subtracting the B-mode images recorded at 70 and 200 kPa, the contrast-to-tissue ratio (CTR) of the native suspension was 3.2dB, whereas the CTR of the filtered suspension was 20 dB. The 17 dB difference is attributed to the threshold behaviour. Well-established ultrasound imaging techniques such as fundamental power modulation imaging could benefit from the pressure-dependent scattering properties of this type of contrast microbubbles.

Medical ultrasound contrast agents (UCAs) have evolved from straight image enhancers to pathophysiological markers and drug delivery vehicles. However, the exact dynamic behavior of the encapsulated bubbles composing UCAs is still not entirely known. In this article, we propose to characterize full populations of UCAs, by looking at the translational effects of ultrasound radiation force on each bubble in a diluted population. The setup involves a sensitive, fully programmable transmitter/receiver and two unconventional, real-time display modes. Such display modes are used to measure the displacements produced by irradiation at frequencies in the range 2-8 MHz and pressures between 150 kPa and 1.5 MPa. The behavior of individual bubbles freely moving in a water tank is clearly observed, and it is shown that it depends on the bubble physical dimensions as well as on the viscoelastic properties of the encapsulation. A new method also is distilled that estimates the viscoelastic properties of bubble encapsulation by fitting the experimental bubble velocities to values simulated by a numerical model based on the modified Herring equation and the Bjerknes force. The fit results are a shear modulus of 18 MPa and a viscosity of 0.23 Pas for a thermoplastic PVC-AN shell. Phospholipid shell elasticity and friction parameter of the experimental contrast agent are estimated as 0.8 N/m and 1 10(-7) kg/s, respectively (shear modulus of 32 MPa and viscosity of 0.19 Pas, assuming 4-nm shell thickness).