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Objective: Local flow dynamics impact atherosclerosis yet are difficult to quantify with conventional ultrasound techniques. This study investigates the performance of ultrasound vector flow imaging (US-VFI) with and without ultrasound contrast agents in the healthy femoral bifurcation., Methods: High-frame-rate ultrasound data with incremental acoustic outputs were acquired in the femoral bifurcations of 20 healthy subjects before (50V) and after contrast injection (2V, 5V and 10V). 2-D blood-velocity profiles were obtained through native blood speckle tracking (BST) and contrast tracking (echo particle image velocimetry [echoPIV]). As a reference, 4-D flow magnetic resonance imaging (4-D flow MRI) was acquired. Contrast-to-background ratio and vector correlation were used to assess the quality of the US-VFI acquisitions. Spatiotemporal velocity profiles were extracted, from which peak velocities (PSV) were compared between the modalities. Furthermore, root-mean-square error analysis was performed., Results: US-VFI was successful in 99% of the cases and optimal VFI quality was established with the 10V echoPIV and BST settings. A good correspondence between 10V echoPIV and BST was found, with a mean PSV difference of -0.5 cm/s (limits of agreement: -14.1-13.2). Both US-VFI techniques compared well with 4-D flow MRI, with a mean PSV difference of 1.4 cm/s (-18.7-21.6) between 10V echoPIV and MRI, and 0.3 cm/s (-23.8-24.4) between BST and MRI. Similar complex flow patterns among all modalities were observed., Conclusion: 2-D blood-flow quantification of femoral bifurcation is feasible with echoPIV and BST. Both modalities showed good agreement compared to 4-D flow MRI. For the femoral tract the administration of contrast was not needed to increase the echogenicity of the blood for optimal image quality., Competing Interests: Conflict of interest This work was supported in part by the Dutch Research Council VORTECS program (17219) and in part by Rijnstate Vriendenfonds (VF19-a10)., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Objective: Ultrasound-triggered bubble-mediated local drug delivery has shown potential to increase therapeutic efficacy and reduce systemic side effects, by loading drugs into the microbubble shell and triggering delivery of the payload on demand using ultrasound. Understanding the behavior of the microbubbles in response to ultrasound is crucial for efficient and controlled release., Methods: In this work, the response of microbubbles with a coating consisting of poly(2-ethyl-butyl cyanoacrylate) (PEBCA) nanoparticles and denatured casein was characterized. High-speed recordings were taken of single microbubbles, in both bright field and fluorescence., Results: The nanoparticle-loaded microbubbles show resonance behavior, but with a large variation in response, revealing a substantial interbubble variation in mechanical shell properties. The probability of shell rupture and the probability of nanoparticle release were found to strongly depend on microbubble size, and the most effective size was inversely proportional to the driving frequency. The probabilities of both rupture and release increased with increasing driving pressure amplitude. Rupture of the microbubble shell occurred after fewer cycles of ultrasound as the driving pressure amplitude or driving frequency was increased., Conclusion: The results highlight the importance of careful selection of the driving frequency, driving pressure amplitude and duration of ultrasound to achieve the most efficient ultrasound-triggered shell rupture and nanoparticle release of protein-and-nanoparticle-stabilized microbubbles., Competing Interests: Conflict of interest The authors declare no competing interests., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Local flow patterns influence stent patency, while blood flow quantification in stents is challenging. The aim of this study was to investigate the feasibility of 2-D blood flow quantification using high-frame-rate, contrast-enhanced ultrasound (HFR-CEUS) and particle image velocimetry (PIV), or echoPIV, in patients with aortoiliac stents. HFR-CEUS measurements were performed at 129 locations in 62 patients. Two-dimensional blood flow velocity fields were obtained using echoPIV. Visual inspection was performed by five observers to evaluate feasibility. The contrast-to-background ratio and average vector correlation were calculated and compared between stented and native vessel segments. Flow quantification with echoPIV was feasible in 128 of 129 locations (99%), with optimal quantification in 40 of 129 locations (31%). Partial quantification was achieved in 88 of 129 locations (68%), where one or multiple limiting issues occurred (not related to the stent) including loss of correlation during systole (57/129), short vessel segments (20/129), loss of contrast during diastole (20/129) and shadow regions (20/129). The contrast-to-background ratio and vector correlation were lower downstream in the imaged blood vessel, independent of the location of the stent. In conclusion, echoPIV was feasible in stents placed in the aortoiliac region, and the stents did not adversely affect flow tracking., Competing Interests: Conflicts of interest disclosure The authors declare no conflicts of interest., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Ultrasound is extensively used in medical imaging, being safe and inexpensive and operating in real time. Its scope of applications has been widely broadened by the use of ultrasound contrast agents (UCAs) in the form of microscopic bubbles coated by a biocompatible shell. Their increased use has motivated a large amount of research to understand and characterize their physical properties as well as their interaction with the ultrasound field and their surrounding environment. Here we review the theoretical models that have been proposed to study and predict the behavior of UCAs. We begin with a brief introduction on the development of UCAs. We then present the basics of free-gas-bubble dynamics upon which UCA modeling is based. We review extensively the linear and non-linear models for shell elasticity and viscosity and present models for non-spherical and asymmetric bubble oscillations, especially in the presence of surrounding walls or tissue. Then, higher-order effects such as microstreaming, shedding and acoustic radiation forces are considered. We conclude this review with promising directions for the modeling and development of novel agents., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

Microbubble ultrasound contrast agents have now been in use for several decades and their safety and efficacy in a wide range of diagnostic applications have been well established. Recent progress in imaging technology is facilitating exciting developments in techniques such as molecular, 3-D and super resolution imaging and new agents are now being developed to meet their specific requirements. In parallel, there have been significant advances in the therapeutic applications of microbubbles, with recent clinical trials demonstrating drug delivery across the blood-brain barrier and into solid tumours. New agents are similarly being tailored toward these applications, including nanoscale microbubble precursors offering superior circulation times and tissue penetration. The development of novel agents does, however, present several challenges, particularly regarding the regulatory framework. This article reviews the developments in agents for diagnostic, therapeutic and "theranostic" applications; novel manufacturing techniques; and the opportunities and challenges for their commercial and clinical translation., Competing Interests: Declaration of Competing Interest TB and SC are full time employees of Bracco Suisse. The other authors are hold full time academic positions and have the following company affiliations: ES is a founder and non-executive director of SonoTarg Ltd.; MB is a founder and chief scientific officer of Respirogen Inc., (Copyright © 2020 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

It has been proposed that monodisperse microbubble ultrasound contrast agents further increase the signal-to-noise ratio of contrast-enhanced ultrasound imaging. Here, the sensitivity of a polydisperse pre-clinical agent was compared experimentally with that of its size- and acoustically sorted derivatives by using narrowband pressure- and frequency-dependent scattering and attenuation measurements. The sorted monodisperse agents had up to a two-orders-of-magnitude increase in sensitivity, that is, in the average scattering cross section per bubble. Moreover, we found, for the first time, that the highly non-linear response of acoustically sorted microbubbles can be exploited to confine scattering and attenuation to the focal region of ultrasound fields used in clinical imaging. This property is a result of minimal pre-focal scattering and attenuation and can be used to minimize shadowing effects in deep tissue imaging. Moreover, it potentially allows for more localized therapy using microbubbles through the spatial control of resonant microbubble oscillations., (Copyright © 2018 World Federation for Ultrasound in Medicine and Biology. Published by Elsevier Inc. All rights reserved.)

For successful in vivo contrast-enhanced ultrasound imaging (CEUS) and ultrasound molecular imaging, detailed knowledge of stability and acoustical properties of the microbubbles is essential. Here, we compare these aspects of lipid-coated microbubbles that have either 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) or 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) as their main lipid; the other components were identical. The microbubbles were investigated in vitro over the frequency range 1-4 MHz at pressures between 10 and 100 kPa, and their response to the applied ultrasound was recorded using ultrahigh-speed imaging (15 Mfps). Relative to DPPC-coated microbubbles, DSPC-coated microbubbles had (i) higher acoustical stability; (ii) higher shell elasticity as derived using the Marmottant model (DSPC: 0.26 ± 0.13 N/m, DPPC: 0.06 ± 0.06 N/m); (iii) pressure amplitudes twice as high at the second harmonic frequency; and (iv) a smaller amount of microbubbles that responded at the subharmonic frequency. Because of their higher acoustical stability and higher non-linear response, DSPC-coated microbubbles may be more suitable for contrast-enhanced ultrasound., (Copyright © 2015 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

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.)

The acoustic characteristics of microbubbles created from vaporized submicron perfluorocarbon droplets with fluorosurfactant coating are examined. Utilizing ultra-high-speed optical imaging, the acoustic response of individual microbubbles to low-intensity diagnostic ultrasound was observed on clinically relevant time scales of hundreds of milliseconds after vaporization. It was found that the vaporized droplets oscillate non-linearly and exhibit a resonant bubble size shift and increased damping relative to uncoated gas bubbles due to the presence of coating material. Unlike the commercially available lipid-coated ultrasound contrast agents, which may exhibit compression-only behavior, vaporized droplets may exhibit expansion-dominated oscillations. It was further observed that the non-linearity of the acoustic response of the bubbles was comparable to that of SonoVue microbubbles. These results suggest that vaporized submicron perfluorocarbon droplets possess the acoustic characteristics necessary for their potential use as ultrasound contrast agents in clinical practice., (Copyright © 2014 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

In this study, we investigated the effect of secondary Bjerknes forces on targeted microbubbles using high-speed optical imaging. We observed that targeted microbubbles attached to an underlying surface and subject to secondary Bjerknes forces deform in the direction of their neighboring bubble, thereby tending toward a prolate shape. The deformation induces an elastic restoring force, causing the bubbles to recoil back to their equilibrium position; typically within 100 μs after low-intensity ultrasound application. The temporal dynamics of the recoil was modeled as a simple mass-spring system, from which a value for the effective spring constant k of the order 10(-3) Nm(-1) was obtained. Moreover, the translational dynamics of interacting targeted microbubbles was predicted by a hydrodynamic point particle model, including a value of the spring stiffness k of the very same order as derived experimentally from the recoiling curves. For higher acoustic pressures, secondary Bjerknes forces rupture the molecular adhesion of the bubbles to the surface. We used this mutual attraction to quantify the binding force between a single biotinylated microbubble and an avidin-coated surface, which was found to be between 0.9 and 2 nanonewtons (nN). The observation of patches of lipids left at the initial binding site suggests that lipid anchors are pulled out of the microbubble shell, rather than biotin molecules unbinding from avidin. Understanding the effect of ultrasound application on targeted microbubbles is crucial for further advances in the realm of molecular imaging., (Copyright © 2013 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

A comparison between phospholipid-coated microbubbles with and without liposomes attached to the microbubble surface was performed using the ultra-high-speed imaging camera (Brandaris 128). We investigated 73 liposome-loaded microbubbles (loaded microbubbles) and 41 microbubbles without liposome loading (unloaded microbubbles) with a diameter ranging from 3-10 μm at frequencies ranging from 0.6-3.8 MHz and acoustic pressures ranging from 5-100 kPa. The experimental data showed nearly the same shell elasticity for the loaded and unloaded bubbles, but the shell viscosity was higher for loaded bubbles compared with unloaded bubbles. For loaded bubbles, a higher pressure threshold for the bubble vibrations was noticed. In addition, an "expansion-only" behavior was observed for up to 69% of the investigated loaded bubbles, which mostly occurred at low acoustic pressures (≤30 kPa). Finally, fluorescence imaging showed heterogeneity of liposome distributions of the loaded bubbles., (Copyright © 2012 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

The dynamics of coated microbubbles was studied in an in vivo model. Biotinylated lipid-coated microbubbles were prepared in-house and were injected into a chick embryo chorioallantoic membrane (CAM) model on the fifth day of incubation. The microbubbles, ranging between 1.0 and 3.5 μm in diameter, were insonified in the frequency range of 4-7 MHz. Two amplitudes of acoustic pressure were applied: 300 kPa and 400 kPa. The fundamental and subharmonic responses were recorded optically with an ultra-fast camera (Brandaris 128) at 20 million frames per second. A subharmonic response was observed for 44% of the studied bubbles. From the data the frequency of the maximum fundamental and subharmonic response was derived for each individual bubble and resulted in the resonance curves of the microbubbles. All the bubbles showed shell (strain) hardening behavior for a higher acoustic pressure. We conclude that the subharmonic oscillations observed in this study belonged to the transmit at resonance (TR) regime., (Copyright © 2012 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

Molecular imaging with ultrasound is a promising noninvasive technique for disease-specific imaging, enabling for instance, the diagnosis of thrombus and inflammation. Selective imaging is performed by using ultrasound contrast agent microbubbles functionalized with ligands, which bind specifically to the target molecules. Here, we investigate in a model system, the influence of adherence at a wall on the dynamics of the microbubbles, in particular, on the frequency of maximum response, by recording the radial response of individual microbubbles as a function of the applied acoustic pressure and frequency. The frequency of maximum response of adherent microbubbles was found to be over 50% lower than for bubbles in the unbounded fluid and over 30% lower than that of a nonadherent bubble in contact with the wall. The change is caused by adhesion of the bubbles to the wall as no influence was found due solely to the presence of the targeting ligands on the bubble dynamics. The shift in the frequency of maximum response may prove to be important for molecular imaging with ultrasound as this application would benefit from an acoustic imaging method to distinguish adherent microbubbles from freely circulating microbubbles., (Copyright © 2011 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

The subharmonic vibration of BR14 (Bracco Research S.A., Geneva, Switzerland) contrast agent microbubbles is investigated within the preferable frequency range for carotid ultrasound imaging (8-12 MHz). The response of the bubbles was recorded optically with an ultra-fast recording camera (Brandaris 128) at three acoustic pressures (50, 100 and 120 kPa). The vibration of the microbubbles was measured as a function of the excitation frequency and its frequency content was determined. Among 390 recordings, 40% showed subharmonic oscillations. It was observed that for smaller microbubbles (diameter < 3 μm) the frequency of the maximum subharmonic response increases for increasing pressures (shell hardening) opposite to what has been reported for larger microbubbles (3 μm < diameter < 15 μm). These findings are well predicted by the model proposed by Marmottant et al. (2005) after including the dilatational shell viscosity of the microbubbles measured by Van der Meer et al. (2007), which indicates a marked shear-thinning behavior of the phospholipid shell., (Copyright © 2011 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

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.)

The influence of the stabilizing phospholipid-coating on the nonlinear dynamics of ultrasound contrast agent microbubbles is investigated. We record the radial dynamics of individual microbubbles with an ultra high-speed camera as a function of both driving pressure and frequency. The viscoelastic shell was found to enhance the nonlinear bubble response at acoustic pressures as low as 10 kPa. For increasing acoustic pressures a decrease of the frequency of maximum response was observed for a distinct class of bubbles, leading to a pronounced skewness of the resonance curve, which we show to be the origin of the "thresholding" behavior (Emmer et al. 2007). For the other bubbles, the frequency of maximum response was found to lie just above the resonance frequency of an uncoated microbubble and to be independent of the applied acoustic pressure. The shell-buckling bubble model (Marmottant et al. 2005), which accounts for buckling and rupture of the shell, captures both cases for a unique set of the shell parameters, the relevant parameter being the phospholipid concentration at the bubble interface., (Copyright © 2010 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

Because the properties of ultrasound contrast agent populations after administration to patients are largely unknown, methods able to study them noninvasively are required. In this study, we acoustically performed a size distribution measurement of the ultrasound contrast agent Definity(®). Single lipid-shelled microbubbles were insonified at 25 MHz, which is considerably higher than their resonance frequency, so that their acoustic responses depended on their geometrical cross sections only. We calculated the size of each microbubble from their measured backscattered pressures. The acoustic size measurements were compared with optical reference size measurements to test their accuracy. Our acoustic sizing method was applied to 88 individual Definity(®) bubbles to derive a size distribution of this agent. The size distribution obtained acoustically showed a mean diameter (2.5 μm) and a standard deviation (0.9 μm) in agreement within 8% with the optical reference measurement. At 25 MHz, this method can be applied to bubble sizes larger than 1.2 μm in diameter. It was observed that similar sized bubbles can give different responses (up to a factor 1.5), probably because of shell differences. These limitations should be taken into account when implementing the method in vivo. This acoustic sizing method has potential for estimating the size distribution of an ultrasound contrast agent noninvasively., (Copyright © 2010 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

Previous optical studies have shown threshold behavior of single-contrast agent microbubbles. Below the acoustic pressure threshold, phospholipid-coated microbubbles with sizes <5.0 mum in diameter oscillate significantly less than above the threshold pressure. Previous studies also revealed an acoustic pressure-dependent attenuation of ultrasound by microbubble contrast agents. In this study, we investigated whether pressure-dependent acoustic behavior may be explained by threshold behavior. For this purpose, pressure-dependent attenuation and scattering of a phospholipid-coated contrast agent were measured. Transmit frequencies between 1.5 and 6.0 MHz and acoustic pressures between 5 and 200 kPa were applied. Unlike the galactose-based contrast agent Levovist, the phospholipid-coated contrast agent BR14 showed a pressure-dependent attenuation. In addition, it was found that filtered suspensions with only microbubbles <3.0 mum in diameter show more pressure-dependent attenuation behavior than native suspensions of phospholipid-coated microbubbles. For the scattering measurements conducted at 3.0 MHz, the native suspension did not show any pressure-dependent behavior. However, the filtered suspension responded highly nonlinearly. Between 30 and 150 kPa, 16 dB additional scattered power was obtained. We concluded that threshold behavior of phospholipid-coated microbubbles results in pressure-dependent attenuation and scattering.

The occurrence of nonspherical oscillations (or surface modes) of coated microbubbles, used as ultrasound contrast agents in medical imaging, is investigated using ultra-high-speed optical imaging. Optical tweezers designed to micromanipulate single bubbles in 3-D are used to trap the bubbles far from any boundary, enabling a controlled study of the nonspherical oscillations of free-floating bubbles. Nonspherical oscillations appear as a parametric instability and display subharmonic behavior: they oscillate at half the forcing frequency, which was fixed at 1.7 MHz in this study. Surface modes are shown to preferentially develop for a bubble radius near the resonance of radial oscillations. In the studied range of acoustic pressures, the growth of surface modes saturates at a level far below bubble breakage. With the definition of a single, dimensionless deformation parameter, the amplitude of nonspherical deformation is quantified as a function of the bubble radius (between 1.5 and 5 microm) and of the acoustic pressure (up to 200 kPa).

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.

Ultrasound contrast agents oscillate approximately linearly up to a certain pressure range where nonlinearity sets in. Nonlinear microbubble oscillations are exploited in ultrasound pulse-echo imaging as this improves the contrast-to-tissue ratio. Here we report the observation of a highly nonlinear response of phospholipid-coated contrast agents at pressures as low as 50 kPa, termed "compression-only" behavior, where the microbubbles only compress, yet hardly expand. Time-resolved bubble dynamics recorded through ultra high-speed imaging revealed that nearly 40% of the coated bubbles show "compression-only" behavior.

Ultrasound contrast agents are now available since a few years and used for diagnostic purposes. Improved diagnostic decisions have been made possible with new imaging methods that are mainly based on the nonlinear properties of gas microbubbles. Since it is well known that contrast agents are destroyed by ultrasound when the acoustic pressure exceeds a threshold, extremely low acoustic pressures were applied to achieve enhanced contrast image quality. However, destruction of contrast microbubbles is not necessarily undesirable, since it is beneficial in, for example, destruction/reperfusion imaging and recently in drug delivery. We investigate in this experimental study the destruction dynamics of a contrast agent consisting of nitrogen bubbles encapsulated in a double polymer/albumin wall shell. This is accomplished using an ultrafast camera Brandaris that operates at a frame rate of 25 MHz and records 128 frames. The measurements were performed with an ultrasound sine burst of 10 cycles at 1.7 MHz. Different acoustic pressures were applied and various microsphere sizes were examined. The results show three different zones depending on the applied pressure and bubble size: these are nondestruction zone, transient zone and destruction zone. The nondestruction zone is reached for either very small microspheres or low mechanical indices (MI) (<0.3). In the destruction zone lie either large microspheres (5 microm or higher) even when irradiated at low MIs or small microspheres (<5 microm) when the MI is above 0.6. The optical observations revealed that the destruction of the microspheres is characterized by shell rupture and gas release. The release of the gas gives rise to new free microbubble that lasts for a few milliseconds and then disappears due to dissolution. In the transient zone, the microspheres are mainly compressed in the first few cycles but no expansion is induced. After intense compressions, the shell fissures and gas escapes in the last cycles of the burst or during a second burst depending on the initial size and MI. These optical recordings are important to investigate contrast bubble destruction and can help in amplifying or minimizing this process. Indeed, bubble disruption remains the basis of most current sensitive methods for detecting perfusion with contrast agents and is an essential component of perfusion quantification with microbubbles, in addition to drug delivery applications and pressure measurements.

Ultrasound (US) in combination with contrast microbubbles has been shown to alter the permeability of cell membranes without affecting cell viability. This permeabilisation feature is used to design new drug delivery systems using US and contrast agents. The underlying mechanisms are still unknown. One hypothesis is that oscillating microbubbles cause cell deformation resulting in enhanced cell membrane permeability. This technical note reveals the interaction between oscillating microbubbles and endothelial cells under a microscope recorded with a fast framing camera at 10 million frames per second. A microbubble expansion of 100% resulted a 2.3-mum displacement of the cell membrane. During the insonification, changes of approximately 15% in the cross-sectional distance of the endothelial cells were observed due to microbubble vibrations. In conclusion, the use of such a camera makes it possible to reveal the mechanisms of interactions between ultrasound, microbubbles and cells.