Acoustic response from adherent targeted contrast agents

In ultrasonic molecular imaging, encapsulated micron-sized gas bubbles are tethered to a blood vessel wall by targeting ligands. A challenging problem is to detect the echoes from adherent microbubbles and distinguish them from echoes from nonadherent agents and tissue. Echoes from adherent contrast agents are observed to include a high amplitude at the fundamental frequency, and significantly different spectral shape compared with free agents (p <0.0003). Mechanisms for the observed acoustical difference and potential techniques to utilize these differences for molecular imaging are proposed.     

Resonance frequencies of lipid-shelled microbubbles in the regime of nonlinear oscillations

Knowledge of resonant frequencies of contrast microbubbles is important for the optimization of ultrasound contrast imaging and therapeutic techniques. To date, however, there are estimates of resonance frequencies of contrast microbubbles only for the regime of linear oscillation. The present paper proposes an approach for evaluating resonance frequencies of contrast agent microbubbles in the regime of nonlinear oscillation. The approach is based on the calculation of the time-averaged oscillation power of the radial bubble oscillation. The proposed procedure was verified for free bubbles in the frequency range 1-4 MHz and then applied to lipid-shelled microbubbles insonified with a single 20-cycle acoustic pulse at two values of the acoustic pressure amplitude, 100 kPa and 200 kPa, and at four frequencies: 1.5, 2.0, 2.5, and 3.0 MHz. It is shown that, as the acoustic pressure amplitude is increased, the resonance frequency of a lipid-shelled microbubble tends to decrease in comparison with its linear resonance frequency. Analysis of existing shell models reveals that models that treat the lipid shell as a linear viscoelastic solid appear may be challenged to provide the observed tendency in the behavior of the resonance frequency at increasing acoustic pressure. The conclusion is drawn that the further development of shell models could be improved by the consideration of nonlinear rheological laws.     

Effect of coupled oscillations on microbubble behavior

Ultrasound contrast agents are encapsulated microbubbles whose nonlinear acoustic scattering signatures have been the foundation of their use in diagnostic imaging. The coupled oscillations of microbubbles along their lines of center are investigated theoretically using radial equations in the monopole approximation and an energy balance, which is obtained for the system. Coupled microbubble pairs of different initial radii are investigated numerically relative to the normal modes for the linearized system. For microbubble pairs of different size bubbles driven below the mode of the smaller bubble and above the mode of the larger bubble, it is shown that oscillations of the smaller agent are affected substantially more by the coupling than those of the larger one. For separation distances of 10 and 500 microns, a difference of approximately 10 dB occurs in the second harmonic output of a 1.0-micron radius agent coupled with a 2.2-micron radius agent forced at 2.0 MHz and 0.3 MPa. The subharmonic spectral peak is shown to decrease approximately 19 dB for the coupling of 1.5- and 2.2-micron radius agents at 10- and 500-micron distances under the same acoustic forcing conditions. These coupling effects on the radiated pressure and its spectral power are highlighted for contrast agent imaging applications.     

Effect of an elastic wall on the dynamics of an encapsulated microbubble: A simulation study

The purpose of the present simulation study is to reveal how confining surfaces with different mechanical properties affect the acoustic response of a contrast agent microbubble. To this end, numerical simulations are carried out for three types of walls: a plastic (OptiCell) wall, an aluminium wall, and a biological tissue. For each wall, the behaviour of contrast microbubbles of three sizes is investigated. The spectral characteristics of the scattered pressure produced by the microbubbles are compared for two cases: the bubble oscillates far away from the wall and the same bubble oscillates in the immediate vicinity of the wall. The results of the simulations allow one to make the following main conclusions. The effect of the OptiCell wall on the acoustic bubble response is stronger than that of the aluminium and tissue walls. Changes in the bubble response near the wall are stronger when bubbles are excited above their resonance frequency. Considering changes in the fundamental and the 2nd harmonic with respect to the peak values of these components at different bubble radii, it is found that the changes are stronger for smaller bubbles and that the changes in the 2nd harmonic are stronger than those in the fundamental. These results allow one to gain an insight into conditions under which the effect of an elastic wall on the acoustic response of a contrast agent microbubble is easier to be detected.     

Ultrasonic scattering cross sections of shell-encapsulated gas bubbles immersed in a viscoelastic liquid: first and second harmonics

The oscillations of gas bubbles, without shell, immersed in viscoelastic liquids and driven by an acoustic wave have been the subject of several investigations. They demonstrate that the viscosity coefficient and the spring constant of the liquid have significant influence on the scattering cross section of the gas bubble. For shell-encapsulated gas bubbles, the investigations have been concentrated to bubbles immersed in a pure viscous liquid. This present work computes the ultrasonic scattering cross section, first and second harmonics, of shell-encapsulated gas bubbles immersed in a viscoelastic liquid. The theoretical model of the bubble oscillation is based on the generalized Rayleigh-Plesset equation of motion of a spherical cavity immersed in a viscoelastic liquid represented by a three-parameter linear Oldroyd model. The scattering cross section is computed for Albunex type of bubble (shell thickness=15 nm, shell shear viscosity=1.77 Pas, shell modulus of rigidity=88.8 MPa) irradiated by a 3.5 MHz ultrasonic pressure wave with an amplitude of 30 kPa. The results demonstrate that encapsulated bubbles respond independently of the surrounding liquid being pure viscous or viscoelastic as long as the surrounding liquid shear viscosity is as low as 10(-3) Pas. Nevertheless, for higher shear viscosities, the bubble responds differently if the surrounding liquid is pure viscous or viscoelastic. In general, the scattering cross sections of first and second harmonics are larger for the viscoelastic liquid.     

Modeling non-spherical oscillations and stability of acoustically driven shelled microbubbles

The oscillation and destruction of microbubbles under ultrasound excitation form the basis of contrast enhanced ultrasound imaging and microbubble assisted drug and gene delivery. A typical microbubble has a size of a few micrometers and consists of a gas core encapsulated by a shell. These bubbles can be driven into surface mode oscillations, which not only contribute to the measured acoustic signal but can lead to bubble destruction. Existing models of surface model oscillations have not considered the effects of a bubble shell. In this study a model was developed to study the surface mode oscillations in shelled bubbles. The effects of shell viscosity and elasticity on the surface mode oscillations were modeled using a Boussinesq-Scriven approach. Simulation was conducted using the model with various bubble sizes and driving acoustic pressures. The occurrence of surface modes and the number of ultrasound cycles needed for the occurrence were calculated. The simulation results show a significant difference between shelled bubbles and shell free bubbles. The shelled bubbles have reduced surface mode amplitudes and a narrower bubble size range within which these modes develop compared to shell free bubbles. The clinical implications were also discussed.     

Ultrasonic bubbles in medicine: influence of the shell

Ultrasound contrast agents consist of microscopically small bubbles encapsulated by an elastic shell. These microbubbles oscillate upon ultrasound insonification, and demonstrate highly nonlinear behavior, ameliorating their detectability. (Potential) medical applications involving the ultrasonic disruption of contrast agent microbubble shells include release-burst imaging, localized drug delivery, and noninvasive blood pressure measurement. To develop and enhance these techniques, predicting the cracking behavior of ultrasound-insonified encapsulated microbubbles has been of importance. In this paper, we explore microbubble behavior in an ultrasound field, with special attention to the influence of the bubble shell. A bubble in a sound field can be considered a forced damped harmonic oscillator. For encapsulated microbubbles, the presence of a shell has to be taken into account. In models, an extra damping parameter and a shell stiffness parameter have been included, assuming that Hooke's Law holds for the bubble shell. At high acoustic amplitudes, disruptive phenomena have been observed, such as microbubble fragmentation and ultrasonic cracking. We analyzed the occurrence of ultrasound contrast agent fragmentation, by simulating the oscillating behavior of encapsulated microbubbles with various sizes in a harmonic acoustic field. Fragmentation occurs exclusively during the collapse phase and occurs if the kinetic energy of the collapsing microbubble is greater than the instantaneous bubble surface energy, provided that surface instabilities have grown big enough to allow for break-up. From our simulations it follows that the Blake critical radius is not a good approximation for a fragmentation threshold. We demonstrated how the phase angle differences between a damped radially oscillating bubble and an incident sound field depend on shell parameters.     

Stability analysis of ultrasound thick-shell contrast agents

The stability of thick shell encapsulated bubbles is studied analytically. 3-D small perturbations are introduced to the spherical oscillations of a contrast agent bubble in response to a sinusoidal acoustic field with different amplitudes of excitation. The equations of the perturbation amplitudes are derived using asymptotic expansions and linear stability analysis is then applied to the resulting differential equations. The stability of the encapsulated microbubbles to nonspherical small perturbations is examined by solving an eigenvalue problem. The approach then identifies the fastest growing perturbations which could lead to the breakup of the encapsulated microbubble or contrast agent.     

Numerical modeling of microbubble backscatter to optimize ultrasound particle image velocimetry imaging: initial studies

We have developed a promising non-invasive ultrasound-based method for performing particle image velocimetry (PIV) in vivo. This method, termed echo PIV, provides multi-component blood velocity data with good (2 ms) temporal resolution. The method takes advantage of the non-linear ultrasound backscatter characteristics of small gas-filled microbubbles (ultrasound contrast) that are seeded into the blood stream. In this study, we use a numerical model to explore potential areas to focus future work in echo PIV.

Ultrasound backscatter from encapsulated microbubbles was modeled using a modified Rayleigh–Plesset equation (Church model, 1995), taking into account the protein/lipid shell layer as a thick, mass-conserving incompressible fluid surrounded by incompressible blood-like fluid. The equation of motion was solved numerically to characterize the fundamental and second harmonic components of the backscattered pressure. Results show a significant advantage in using the second harmonic component for echo PIV, especially for small bubble sizes less than 3 μm in diameter at 2.2 MHz frequency. The effect of the shell thickness ranging from 10 to 500 nm on the vibration amplitude of the bubble was examined and it is shown that the presence of the shell requires mechanical index (MI)>0.2 of incident pressure amplitude to improve bubble detectability. Analysis of the effect of pulse length shows a tradeoff between axial resolution (short pulse length) and bubble detectability (longer pulse length) will most likely be required. The effect of varying MI between 0.1 and 0.6 was also studied at a center frequency of 2.2 MHz and the results indicate that the resonance of the second harmonic is maximized for bubbles with diameter of approximately 2.75 μm. Bubble non-linearities at MI>0.2 induced a resonant frequency shift away from the integer multiple of the incident frequency in the second harmonic backscatter. For a given bubble size, there is a combination of optimal incident frequency and mechanical index range that maximizes the ratio of the second harmonic compared to the fundamental. This resonant frequency decreases with increasing bubble radius. Further, a narrow bandwidth pulse is shown to increase signal strength. Both of these effects may cause conflict with factors governing spatial resolution. Optimization of the incident frequency, microbubble size and mechanical index to enhance bubble detectability will depend on the particular clinical application. These theoretical predictions provide further understanding of the physics behind our echo PIV technique, and should be useful for guiding the design of echo PIV systems.     

The dependence of ultrasound contrast agents backscatter on acoustic pressure: theory versus experiment

Experimental investigations have not fully explored the interaction between ultrasound beams and microbubble contrast agents. Moreover theoretical investigations have not solved the problem of the microbubble oscillation. A simple in-vitro system based on a commercial scanner (ATL UM9) was used to insonate (3 MHz transmission) diluted contrast suspensions of Definity and Quantison at different acoustic pressures (0.27-1.52 MPa). The experimental data were referred to a blood mimicking fluid in order to extract an estimate of their scattering cross-section. The results were compared with the solutions of the three main bubble oscillatidn models, Rayleigh-Plesset, Herring and Gilmore. Non-linear solutions of the above models were produced numerically using the Mathematica Package Software. The experiments showed that both agents provided a linear increase in scattering cross-section with increasing acoustic pressure. The thick shelled Quantison provided an increasing number of scatterers with increasing acoustic pressure, which proved that free bubbles leaked out of the shell. At high acoustic pressures both Quantison and Definity scattering cross-sections were almost identical, and were probably that of a free bubble. The Rayleigh-Plesset model provided a scattering cross-section almost independent of acoustic pressure. On the contrary the scattering cross-sections calculated by the Herring and Gilmore models solutions displayed a definite dependence on acoustic pressure of an order higher than one, which is slightly higher than the order of dependence exhibited by the experimental data. However, the increase of the experimentally measured scattering cross-section with acoustic pressure was sharper than the calculated one by the above two models. This is most probably due to the fact that the models simulated damped and not free bubble oscillations. In conclusion the Rayleigh-Plesset model was inadequate in describing the bubble oscillations even at small diagnostic acoustic pressures. The Herring and Gilmore models could simulate the dependence of the scattering cross-section of encapsulated microbubbles on acoustic pressure. However the contribution of free bubble oscillations has still to be modelled.     

Difference frequency and its harmonic emitted by microbubbles under dual frequency excitation

Vibro-acoustography is an elasticity imaging method that uses two ultrasound beams of slightly different frequency to excite an object and detects the resulting acoustic emission (AE) at the difference frequency. This method is especially sensitive to bubbles due to their nonlinearity. This study explores the harmonic acoustic emission (HAE) at twice the difference frequency emitted from bubbles. A perturbation method based on the dynamic bubble equation is used to derive the AE and HAE from a single bubble excited by dual frequency waves. Simulation shows that HAE is generated only by microbubbles whose resonant frequencies match the incident ultrasound frequencies. In contrast, AE is more sensitive to resonance at the difference frequency, which is relevant to sub-millimeter bubbles. This finding was confirmed by experiments where HAE was produced from Optison microbubbles, but not from larger air bubbles which are off resonance at the incident ultrasound frequency. In conclusion, harmonic acoustic emission is present for microbubbles. It is very sensitive to the size of the bubble and may be used for selective detection of microbubbles.     

Physical forces exerted by microbubbles on a surface in a traveling wave field

The effect of a wave with a varying traveling component on the bubble activity as well as the physical force generated by microbubbles on a surface has been studied. The acoustic emission from a collection of bubbles is measured in a 928 kHz sound field. Particle removal tests on a surface, which actually measures the applied physical force by the bubbles on that surface, indicate a very strong dependence on the angle of incidence. In other words, when the traveling wave component is maximized, the average physical force applied by microbubbles reaches a maximum. Almost complete particle removal for 78 nm silica particles was obtained for a traveling wave, while particle removal efficiency was reduced to only a few percent when a standing wave was applied. This increase in particle removal for a traveling wave is probably caused by a decrease in bubble trapping at nodes and antinodes in a standing wave field.     

Influence of wake bubbles on optical impulse scattering echo

The model of the optical impulse scattering echo based on Mie theory and backscattering properties of bubble populations is presented. The experimental results of optical impulse scattering echo for a ship in the ocean is in agreement with the simulated experiment. The wake bubbles significantly influence the laser scattering echo process in the ocean. When the testing system is outside the wake, the distinct scattering echoes of interface between wake bubbles and water are received. When the testing system is inside the wake, the intensity of the echo attenuates with the increased scattering of wake bubbles. These phenomena of laser echoes in different periods are analyzed.     

Optimal Acoustic Attenuation of Weakly Compressible Media Permeated with Air Bubbles

Based on fuzzy logic （FL） and genetic algorithm （GA）, we present an optimization method to obtain the optimal acoustic attenuation of a longitudinal acoustic wave propagating in a weakly compressible medium permeated with air bubbles. In the optimization, the parameters of the size distribution of bubbles in the medium are optimized for providing uniformly high acoustic attenuation in the frequency band of interest. Compared with other traditional optimization methods, the unique advantage of the present method is that it can locate the global optimum quickly and effectively in need of knowing the mathematical model precisely. As illustrated by a numerical simulation, the method is effective and essential in enhancing the acoustic attenuation of such a medium in an optimal manner. The bubbly medium with optimized structural parameters can effectively attenuate longitudinal waves at intermediate frequencies with an acoustic attenuation approximating a constant value of lO（dB/cm）. Such bubbly media with optimal acoustic attenuations may be applied to design acoustic absorbent by controlling broader attenuation band and higher efficiency.     

The ultrasonic weak short-pulse responses of microbubbles based on a two-frequency approximation

The ultrasonic short-pulse responses of microbubbles are of interest in cavitation, transient responses, and contrast imaging. We extend the two-frequency analytic solutions of Newhouse and Shankar [J. Acoust. Soc. Am. 75, 1473-1477 (1984)] to approximate the short-pulse responses of microbubbles in a low-amplitude field. Based on their results, there is an expected component near dc in the spectrum of bubble echoes excited by a short pulse. Here this component is named the low-frequency response, and its theoretical properties are verified experimentally. Including the fundamental and second-harmonic components, the weak short-pulse responses of microbubbles include three types of response. Our work has determined the constraint conditions under which this approximated solution can be used to analyze these short-pulse responses. This paper also provides the amplitude and spectral properties of these responses. The low-frequency response has a special bandwidth-dependent property and has potential applications in imaging and bubble sizing.     

Real-Time Measurements and Modelling on Dynamic Behaviour of SonoVue Bubbles Based on Light Scattering Technology

The dynamic behaviour of SonoVue microbubbles, a new generation ultrasound contrast agent, is investigated in real time with light scattering method. Highly diluted SonoVue microbubbles are injected into a diluted gel made of xanthan gum and water. The responses of individual SonoVue bubbles to driven ultrasound pulses are measured. Both linear and nonlinear bubble oscillations are observed and the results suggest that SonoVue microbubbles can generate strong nonlinear responses. By fitting the experimental data of individual bubble responses with Sarkar＇s model, the shell coating parameter of the bubbles and dilatational viscosity is estimated to be 7.0 nm-s-Pa.     

Modelling of nonlinear effects and the response of ultrasound contrast micro bubbles: simulation and experiment

The propagation of diagnostic ultrasonic imaging pulses in tissue and their interaction with contrast micro bubbles is a very complex physical process, which we assumed to be separable into three stages: pulse propagation in tissue, the interaction of the pulse with the contrast bubble, and the propagation of the scattered echo. The model driven approach is used to gain better knowledge of the complex processes involved. A simplified way of field simulation is chosen due to the complexity of the task and the necessity to estimate comparative contributions of each component of the process. Simulations are targeted at myocardial perfusion estimation. A modified method for spatial superposition of attenuated waves enables simulations of low intensity pulse pressure fields from weakly focused transducers in a nonlinear, attenuating, and liquid-like biological medium. These assumptions enable the use of quasi-linear calculations of the acoustic field. The simulations of acoustic bubble response are carried out with the Rayleigh-Plesset equation with the addition of radiation damping. Theoretical simulations with synthesised and experimentally sampled pulses show that the interaction of the excitation pulses with the contrast bubbles is the main cause of nonlinear scattering, and a 2-3 dB increase of second harmonic amplitude depends on nonlinear distortions of the incident pulse.     

两种气泡混合的声空化

将非线性声波方程和改进的Rayleigh-Plesset方程联立可以描述空化环境中的声场及相应的气泡动力学特征. 用时域有限差分方法模拟了圆柱形容器内两种气泡相互混合时的空化情况. 在烧杯内的稳态背景声场形成过程中, 瓶壁耗散吸收扮演了重要的角色. 在稳态背景声场的基础上, 分析了混合气泡与声场的相互作用、气泡之间的相互作用、混合情况下的频谱特性. 结果表明: 两种气泡平衡半径都不太大时, 气泡与声场的相互作用不强, 声场及气泡的行为也比较规律; 相反, 当其中一种气泡平衡半径相对比较大时, 声场与气泡具有较强的非线性相互作用, 声场及气泡的行为表现出复杂的特性.     

Modeling of nonlinear viscous stress in encapsulating shells of lipid-coated contrast agent microbubbles

A general theoretical approach to the development of zero-thickness encapsulation models for contrast microbubbles is proposed. The approach describes a procedure that allows one to recast available rheological laws from the bulk form to a surface form which is used in a modified Rayleigh-Plesset equation governing the radial dynamics of a contrast microbubble. By the use of the proposed procedure, the testing of different rheological laws for encapsulation can be carried out. Challenges of existing shell models for lipid-encapsulated microbubbles, such as the dependence of shell parameters on the initial bubble radius and the “compression-only” behavior, are discussed. Analysis of the rheological behavior of lipid encapsulation is made by using experimental radius-time curves for lipid-coated microbubbles with radii in the range 1.2-2.5 μm. The curves were acquired for a research phospholipid-coated contrast agent insonified with a 20 cycle, 3.0 MHz, 100 kPa acoustic pulse. The fitting of the experimental data by a model which treats the shell as a viscoelastic solid gives the values of the shell surface viscosity increasing from 0.30 × 10⁻⁸ kg/s to 2.63 × 10⁻⁸ kg/s for the range of bubble radii, indicated above. The shell surface elastic modulus increases from 0.054 N/m to 0.37 N/m. It is proposed that this increase may be a result of the lipid coating possessing the properties of both a shear-thinning and a strain-softening material. We hypothesize that these complicated rheological properties do not allow the existing shell models to satisfactorily describe the dynamics of lipid encapsulation. In the existing shell models, the viscous and the elastic shell terms have the linear form which assumes that the viscous and the elastic stresses acting inside the lipid shell are proportional to the shell shear rate and the shell strain, respectively, with constant coefficients of proportionality. The analysis performed in the present paper suggests that a more general, nonlinear theory may be more appropriate. It is shown that the use of the nonlinear theory for shell viscosity allows one to model the “compression-only” behavior. As an example, the results of the simulation for a 2.03 μm radius bubble insonified with a 6 cycle, 1.8 MHz, 100 kPa acoustic pulse are given. These parameters correspond to the acoustic conditions under which the “compression-only” behavior was observed by de Jong et al. [Ultrasound Med. Biol. 33 (2007) 653-656]. It is also shown that the use of the Cross law for the modeling of the shear-thinning behavior of shell viscosity reduces the variance of experimentally estimated values of the shell viscosity and its dependence on the initial bubble radius.     

Impact of acoustic pressure on ambient pressure estimation using ultrasound contrast agent

Local blood pressure measurements provide important information on the state of health of organs in the body and can be used to diagnose diseases in the heart, lungs, and kidneys. This paper presents an approach for investigating the ambient pressure sensitivity of a contrast agent using diagnostic ultrasound. The experimental setup resembles a realistic clinical setup utilizing a single array transducer for transmit and receive. The ambient pressure sensitivity of SonoVue (Bracco, Milano, Italy) was measured twice using two different acoustic driving pressures, which were selected based on a preliminary experiment. To compensate for variations in bubble response and to make the estimates more robust, the relation between the energy of the subharmonic and the fundamental component was chosen as a measure over the subharmonic peak amplitude. The preliminary study revealed the growth stage of the subharmonic component to occur at acoustic driving pressures between 300 and 500 kPa. Based on this, the pressure sensitivity was investigated using a driving pressure of 485 and 500 kPa. At 485 kPa, a linear pressure sensitivity of 0.42 dB/kPa was found having a linear correlation coefficient of 0.94. The second measurement series at 485 kPa showed a sensitivity of 0.41 dB/kPa with a correlation coefficient of 0.89. Based on the measurements at 500 kPa, this acoustic driving pressure was concluded to be too high causing the bubbles to be destroyed. The pressure sensitivity for these two measurement series were 0.42 and 0.25 dB/kPa with linear correlation coefficients of 0.98 and 0.93, respectively.