Radial modulation of microbubbles for ultrasound contrast imaging

Over the past few years, extensive research has been carried out in the field of ultrasound contrast imaging. In addition to the development of new types of ultrasound contrast agents, various imaging methods dedicated to contrast agents have been introduced, and some of them are now commercially available. In this study, we present results of an imaging technique that is capable of detecting echoes from microbubbles and eliminating those emanating from nonoscillating structures (tissue), thereby enhancing contrast imaging. The method is based on mixing a low frequency (LF) modulator signal and a high frequency (HF) imaging signal to effectively modulate the size of the contrast microbubble through its volumetric oscillations using the LF signal and to probe the radial motion using the HF imaging signal. To evaluate the performances and limitations of the method, high-speed optical observations and acoustic measurements were carried out on soft-shelled microbubbles. The results showed that, by incorporating the modulator signal, the bubbles respond differently compared to the HF excitation alone. The decorrelation between the signals obtained at the compression and expansion phase of the modulator signal is significantly high to be used as a parameter to detect contrast microbubbles and discriminate them from tissue. The echo received from a solid reflector shows identical responses during the compression and rarefaction phase of the LF signal. In conclusion, these results demonstrate the feasibility of this fully linear approach for improving the contrast detection.     

Optical and acoustic detection of laser-generated microbubbles in single cells

Acoustically monitored laser-induced optical breakdown (LIOB) has potential as an important tool to diagnose and treat living cells. Laser-induced intracellular microbubbles are readily detectable using high-frequency ultrasound, and LIOB can be controlled to operate within two distinct regimes. In the nondestructive regime, a single, short-lived bubble can be generated within a cell, without affecting its immediate viability. In the destructive regime, the induced photodisruption quickly can kill a targeted cell. To generate and monitor this range of bioeffects in real time, we have developed a system integrating an ultrafast laser source with optical and acoustic microscopy. Experiments were performed on monolayers of Chinese hamster ovary (CHO) cells. A 793 nm, 100 fs laser pulsed at 3.8 kHz was tightly focused within each cell to produce the photodisruption, and a 50 MHz ultrasonic transducer monitored the resultant bubble via continuous pulse-echo recordings. Photodisruption was also observed using bright field microscopy, and cell viability was assessed following laser exposure with a trypan blue assay. By controlling laser pulse fluence and exposure duration, either nondestructive or destructive LIOB could be produced. The intracellular position of the laser focus was also varied to demonstrate that cell viability was affected by the specific location of material breakdown.     

Dynamics and fragmentation of thick-shelled microbubbles

Localized delivery could decrease the systemic side effects of toxic chemotherapy drugs. The unique delivery agents we examine consist of microbubbles with an outer lipid coating, an oil layer, and a perfluorobutane gas core. These structures are 0.5-12 /spl mu/m in radius at rest. Oil layers of these acoustically active lipospheres (AALs) range from 0.3-1.5 /spl mu/m in thickness and thus the agents can carry a large payload compared to nano-scale drug delivery systems. We show that triacetin-based drug-delivery vehicles can be fragmented using ultrasound. Compared with a lipid-shelled contrast agent, the expansion of the drug-delivery vehicle within the first cycle is similar, and a subharmonic component is demonstrated at an equivalent radius, frequency, and driving pressure. For the experimental conditions explored here, the pulse length required for destruction of the drug-delivery vehicle is significantly greater, with at least five cycles required, compared with one cycle for the contrast agent. For the drug-delivery vehicle, the observed destruction mechanism varies with the initial radius, with microbubbles smaller than resonance size undergoing a symmetric collapse and producing a set of small, equal-sized fragments. Between resonance size and twice resonance size, surface waves become visible, and the oscillations become asymmetrical. For agents larger than twice the resonance radius, the destruction mechanism changes to a pinch-off, with one fragment containing a large fraction of the original volume.     

Detecting microbubbles in transformer oil

The growth of microbubbles under the action of a negative pressure wave in transformer oil (TO) has been studied. It is established that microbubbles in TO grow to a detectable size of 100–200 μm. According to the results of calculations, these bubbles can develop from the nuclei of micron and submicron dimensions.     

Magnetophoretic velocity modulation mass analysis of a single microparticle in an atmosphere

A new principle of the magnetophoretic velocity modulation mass analysis of microparticles, which can determine simultaneously the mass and magnetic susceptibility of a single microparticle, has been proposed, and the measurement system was constructed by applying a magnetophoretic force on a falling microparticle through a magnetic field gradient in an atmosphere. A polystyrene microparticle as a test particle adsorbed on a glass plate was selectively knocked off by a pulsed Nd:YAG laser impact into a narrow gap of pole pieces of permanent magnets having a magnetic field gradient with a maximum intensity of 850 T2 m(-1). The falling particle was irradiated by a He-Ne laser, and the scattered light was detected through a slit array mask as a function of time. A bundle of spiked signals of scattered light intensity was analyzed to obtain velocities, which gave acceleration and deceleration of the falling particle. On the basis of the equation of motion under the magnetic field gradient, the mass and magnetic susceptibility of the test particle were reasonably determined.     

The influence of blood on targeted microbubbles

The ability to successfully target the delivery of drugs and other therapeutic molecules has been a key goal of biomedical research for many decades. Despite highly promising in vitro results, however, successful translation of targeted drug delivery into clinical use has been extremely limited. This study investigates the significance of the characteristics of whole blood, which are rarely accounted for in vitro assays, as a possible explanation for the poor correlation between in vitro and in vivo experiments. It is shown using two separate model systems employing either biochemical or magnetic targeting that blood causes a substantial reduction in targeting efficiency relative to saline under the same flow conditions. This finding has important implications for the design of targeted drug delivery systems and the assays used in their development.     

Multiple emulsion microbubbles for ultrasound imaging

In order to improve the sensitivity of ultrasound imaging, the contrast agents, a powerful non-invasive and real-time medical imaging technique, are used. However, air or N₂ or perfluorocarbon only encapsulated microbubbles which are currently used have lower efficiency and short imaging time. So the novel contrast agents with a higher efficiency are required. To achieve this objective, the strategy that we have explored involves the use of superparamagnetic iron oxide (SPIO) Fe₃O₄ nanoparticles multilayer emulsion microbubbles. This multilayer structure consists of three layers. The core is poly-d, l-lactide (PLA) encapsulated N₂ nanobubble with the SPIO nanoparticles forming oil-in-water (W/O) layer. The outermost is water-in-oil-in-water ((W/O)/W) emulsion layer with PVA solution. Herein we describe the synthesis and characterization of ultrasound imaging microstructure with an overall diameter of around 2μm-8μm. On the one hand, the stable gas encapsulated microstructure can provide a high scattering intensity resulting in high echogenicity, On the other hand, SPIO nanoparticles have shown the potential of high-resolution sonography. So the multiple emulsion microbubbles with SPIO can have double action to enhance the ultrasound imaging. Besides, because SPIO can also serve as magnetic resonance imaging (MRI) contrast agents, such microstructure may be useful for multimodality imaging studies in ultrasound imaging and MRI.     

Review of shell models for contrast agent microbubbles

Micrometer-scale encapsulated gas bubbles, known as ultrasound contrast agents, are used in ultrasound medical diagnostics for enhancing blood-tissue contrast during an ultrasonic examination. They are also employed in therapy as an activator of drug incorporation or extravasation. Adequate modeling of the effect of encapsulation is of primary importance because it is the encapsulating shell that determines many of the functional properties of contrast agents. In this review, existing approaches to the modeling of the radial motion of an encapsulated bubble are discussed and comparative analysis of available shell models is conducted. The capabilities of the shell models are evaluated in the context of recent experimental observations, such as compression-only behavior and the dependence of shell material properties on initial bubble radius. It is shown that for early shell models, the main problem is that the behavior of encapsulation is described by linear elastic and viscous laws, whereas recent experimental data attest to complicated rheological properties inherent in shell materials. Currently, a trend toward models involving nonlinear laws for shell elasticity and viscosity is observed. In particular, nonlinear models have been proposed that allow one to reproduce compression-only behavior. However, the problem of the radius dependence of shell material parameters remains unsolved.     

Gas-storage ice grown from water containing microbubbles

Gas-storage ice is the ice that contains various functional gas pores, and can be produced by freezing water in which a desired gas is dissolved. In this conventional method, however, the gas content in the ice is limited by the gas solubility in water. To overcome this limitation, we developed a method to produce gas-storage ice from water in which microbubbles of a desired gas are dispersed, and then obtained images of the structural features of pores formed in the ice prepared using this microbubble method. The images clarified the interaction between a microbubble arriving at an ice–water interface and an existing pore already formed in the ice. The air content in ice prepared using the microbubble method was higher than that prepared using the conventional method, and was about three times higher than the solubility of air in water.     

Ultrasound-induced gas release from contrast agent microbubbles

We investigated gas release from two hardshelled ultrasound contrast agents by subjecting them to high-mechanical index (MI) ultrasound and simultaneously capturing high-speed photographs. At an insonifying frequency of 1.7 MHz, a larger percentage of contrast bubbles is seen to crack than at 0.5 MHz. Most of the released gas bubbles have equilibrium diameters between 1.25 and 1.75 microm. Their disappearance was observed optically. Free gas bubbles have equilibrium diameters smaller than the bubbles from which they have been released. Coalescence may account for the long dissolution times acoustically observed and published in previous studies. After sonic cracking, the cracked bubbles stay acoustically active.     

Gradient polymer-disposed liquid crystal single layer of large nematic droplets for modulation of laser light

The light modulating ability of gradient polymer-disposed liquid crystal (PDLC) single layer of large droplets formed by nematic E7 in UV-cured polymer NOA65 is studied. Operating at relatively low voltages, such PDLC film with a of thickness 10-25?μm and droplet size up to 50?μm exhibits a good contrast ratio and is capable of producing a large phase shift for the propagating coherent light. For a linearly polarized He-Ne laser (λ=633?nm), an electrically commanded phase shift as large as π/2 can be obtained by the large-droplet region of the film. The electrically produced phase shift and its spatial profile controlled by the thickness of the gradient PDLC single layers of large nematic droplets can be useful for tunable spatial light modulators and other devices for active control of laser light.     

陶瓷微孔膜管制造微气泡的研究

气浮工艺中气泡的大小是衡量气泡制造技术的关键.研究了陶瓷微孔膜管产生微气泡的条件,并对两种不同材质的微孔膜管产生微气泡的结果进行了比较,从气泡形成机理上分析了影响气泡粒径分布的因素.实验采用静态显微摄像技术和图像分析系统时气泡粒径分布进行了表征.实验结果表明,利用陶瓷微孔膜管产生的气泡粒径主要集中在15～50 μm之间,平均粒径在25.7～44.2 μm之间.微孔膜管的孔径大小、表面性能,气体流量,剪切流流速,液体表面张力,黏度是影响气泡粒径分布的主要因素.     

Reflection from bound microbubbles at high ultrasound frequencies

Targeted contrast agents and ultrasound imaging are now used in combination for the assessment and tracking of biomarkers in animal models in vivo. These applications have triggered interest in the understanding and prediction of the ultrasound echoes from contrast agents attached to cells. This study describes the reflection enhancement due to microbubbles bound on a gelatin surface. The reflection enhancement was measured using ultrasound pulses at high frequency (40 MHz) and low pressure (38 kPa peak-negative pressure) allowing a linear approximation to be applied. The observed reflection coefficient increased with the number of microbubbles, until reaching saturation at 0.9 when the surface coverage fraction was 35%. A multiple scattering model assuming that the targeted microbubbles are confined within an infinitesimally thin layer appeared suitable in predicting the reflection coefficient even at very high surface densities. These results could permit the optimization of the sensitivity of high frequency ultrasound to targeted contrast agents.     

Removal of residual pesticides in vegetables using ozone microbubbles

The removal of fenitrothion (FT) pesticide residues from vegetables by immersion in ozone-microbubbled solution was demonstrated. FT-treated lettuce, cherry tomatoes, and strawberries were immersed in ozone-microbubbled, ozone-millibubbled, and dechlorinated water. After that the percentage of residual FT in the vegetables was determined. Residual FT was efficiently removed from lettuce by immersing it in ozone-microbubbled solution containing more than 1.0 ppm dissolved ozone, or continuously generated ozone-microbubbled solution containing 2.0 ppm dissolved ozone. Similarly, for cherry tomatoes and strawberries, the continuously generated ozone-microbubbled solution containing 2.0 ppm dissolved ozone was highly effective. These results showed that ozone microbubbles effectively removed residual pesticides not only from leafy vegetables but also from fruity vegetables.     

Characterization of cell membrane response to ultrasound activated microbubbles

Contrast agents for ultrasound imaging, composed of tiny gas microbubbles, have become a reality in clinical routine. They are extensively used in radiology for detection and characterization of various tumors and in cardiology for left ventricular opacification. Recent experimental studies showed that ultrasound waves in combination with contrast agent microbubbles increase transiently cell membrane permeability in a process known as sonoporation. This effect is thought to allow foreign molecules to enter the cell. In that context, we explored the cell membrane's responses to microbubbles' oscillations as the mechanism is not completely understood. Breast cancer cell line in combination with contrast microbubbles were used. Ultrasound was applied using a transducer of 1 MHz center frequency transmitting a 10-cycle burst of different acoustic pressures repeated every 100 mus. Patch-clamp technique in whole cell configuration was used to explore transmembrane ion exchange through the variations in membrane potential. To characterize the activated ion channels, the variations of the intracellular calcium (Ca(2+)) concentration were explored using a fluorescent marker. The results revealed that ultrasound stimulation induces a rapid hyperpolarization of cell membrane potential when the microbubble is in direct contact with the cell, but the potential returned to its initial value when ultrasound stimulation stopped. The change in cell membrane potential indicates the activation of specific ion channels and depends on the quality of microbubble adhesion to the cell membrane. Microbubbles were shown to induce a mechanical stretch activating BKca channels. Simultaneous Ca(2+) measurements indicate a slow and progressive Ca(2+) increase that is likely a consequence of BKca channels opening not a cause. These results demonstrate that microbubbles' oscillations under ultrasound activation entail modulation of cellular function and signaling by t- riggering the modulation of ionic transports through the cell membrane. Cells response to the mechanical stretch caused by gentle microbubble oscillations is characterized by the opening of BKca stretch channels and a Ca(2+) flux, which might potentially trigger other cellular responses responsible for membrane sonopermeabilization.     

Submolecular resolution viscoelastic imaging of a poly(p-toluene-sulfonate) single crystal using force modulation microscopy

Submolecular resolution viscoelastic imaging of a poly(2,4-hexadiyne-1,6-diyl bis(p-toluene-sulfonate)) single crystal was achieved using force modulation microscopy under ambient conditions. The elastic image clearly visualized the structure of p-toluene-sulfonate side chains. The viscotic image visualized that the phase delay on the main chains was smallest, while it became largest on the toluene rings in the side chains. The result is considered to be closely related to the molecular dynamics of the crystal.     

水中微气泡光散射偏振特性的研究

在Mie散射理论的基础上，建立气泡光散射模型，计算水中微小气泡与固体微粒在不同的粒径参数和散射角条件下的光散射偏振特性，并对两者进行比较．结果表明，水中气泡及微粒对入射光散射后偏振状态的改变十分复杂．总体上气泡的退偏振效应比固体微粒强25％，微小气泡与固体微粒对偏振状态的影响在粒径域和散射角上具有选择性．     

Preparation and antitumor activity of bFGF-mediated active targeting doxorubicin microbubbles

Characterization and antitumor activity of basic fibroblast growth factor-mediated active targeting doxorubicin microbubbles (bFGF-DOX-MB) were investigated. Pluronic F68 with chemical conjugation of doxorubicin (DOX-P) and peptide KRTGQYKLC-conjugated DSPE–PEG2000 were prepared. bFGF-DOX-MB had a normal distribution of particle size, with average particle size of 2.7 μm. Using A549 mouse model, bFGF-DOX-MB combined ultrasound showed the best inhibition effect on tumor volume growth among all the test groups. Similar conclusion was obtained from experimental measurements of tumor weight change and blood cell count. From the results, chemotherapeutic drug inhibition on tumor growth could be enhanced by local ultrasound combined with active targeting bFGF-DOX-MB, which might provide a potential application for ultrasound-mediated chemotherapy.