Chee Hau Leow

Flow Velocity Mapping Using Contrast Enhanced High-Frame-Rate Plane Wave Ultrasound and Image Tracking: Methods and Initial in Vitro and in Vivo Evaluation

Ultrasound imaging is the most widely used method for visualising and quantifying blood flow in medical practice, but existing techniques have various limitations in terms of imaging sensitivity, field of view, flow angle dependence, and imaging depth. In this study, we developed an ultrasound imaging velocimetry approach capable of visualising and quantifying dynamic flow, by combining high-frame-rate plane wave ultrasound imaging, microbubble contrast agents, pulse inversion contrast imaging and speckle image tracking algorithms. The system was initially evaluated in vitro on both straight and carotid-mimicking vessels with steady and pulsatile flows and in vivo in the rabbit aorta. Colour and spectral Doppler measurements were also made. Initial flow mapping results were compared with theoretical prediction and reference Doppler measurements and indicate the potential of the new system as a highly sensitive, accurate, angle-independent and full field-of-view velocity mapping tool capable of tracking and quantifying fast and dynamic flows.

Effects of microchannel confinement on acoustic vaporisation of ultrasound phase change contrast agents

The sub-micron phase change contrast agent (PCCA) composed of a perfluorocarbon liquid core can be activated into gaseous state and form stable echogenic microbubbles for contrast-enhanced ultrasound imaging. It has shown great promise in imaging microvasculature, tumour microenvironment, and cancer cells. Although PCCAs have been extensively studied for different diagnostic and therapeutic applications, the effect of biologically geometrical confinement on the acoustic vaporisation of PCCAs is still not clear. We have investigated the difference in PCCA-produced ultrasound contrast enhancement after acoustic activation with and without a microvessel confinement on a microchannel phantom. The experimental results indicated more than one-order of magnitude less acoustic vaporisation in a microchannel than that in a free environment taking into account the attenuation effect of the vessel on the microbubble scattering. This may provide an improved understanding in the applications of PCCAs in vivo.

MICROBUBBLE VOID IMAGING: A NON-INVASIVE TECHNIQUE FOR FLOW VISUALISATION AND QUANTIFICATION OF MIXING IN LARGE VESSELS USING PLANE WAVE ULTRASOUND AND CONTROLLED MICROBUBBLE CONTRAST AGENT DESTRUCTION

There is increasing recognition of the influence of the flow field on the physiology of blood vessels and their development of pathology. Preliminary work is reported on a novel non-invasive technique, microbubble void imaging, which is based on ultrasound and controlled destruction of microbubble contrast agents, permitting flow visualisation and quantification of flow-induced mixing in large vessels. The generation of microbubble voids can be controlled both spatially and temporally using ultrasound parameters within the safety limits. Three different model vessel geometries-straight, planar-curved and helical-with known effects on the flow field and mixing were chosen to evaluate the technique. A high-frame-rate ultrasound system with plane wave transmission was used to acquire the contrast-enhanced ultrasound images, and an entropy measure was calculated to quantify mixing. The experimental results were cross-compared between the different geometries and with computational fluid dynamics. The results indicated that the technique is able to quantify the degree of mixing within the different configurations, with a helical geometry generating the greatest mixing, and a straight geometry, the lowest. There is a high level of concordance between the computational fluid dynamics and experimental results. The technique could also serve as a flow visualisation tool.

Spatio-Temporal Flow and Wall Shear Stress Mapping Based on Incoherent Ensemble-Correlation of Ultrafast Contrast Enhanced Ultrasound Images

In this study, a technique for high-frame-rate ultrasound imaging velocimetry (UIV) is extended first to provide more robust quantitative flow velocity mapping using ensemble correlation of images without coherent compounding, and second to generate spatio-temporal wall shear stress (WSS) distribution. A simulation model, which couples the ultrasound simulator with analytical flow solution, was implemented to evaluate its accuracy. It is shown that the proposed approach can reduce errors in velocity estimation by up to 10-fold in comparison with the coherent correlation approach. Mean errors (ME) of 3.2% and 8.6% were estimated under a steady flow condition, while 3.0% and 10.6% were found under a pulsatile condition for the velocity and wall shear rate (WSR) measurement, respectively. Appropriate filter parameters were selected to constrain the velocity profiles before WSR estimations and the effects of incorrect wall tracking were quantified under a controlled environment. Although accurate wall tracking is found to be critical in WSR measurement (as a 200 µm deviation from the wall may yield up to a 60% error), this can be mitigated by HFR imaging (of up to 10 kHz) with contrast agents, which allow for improved differentiation of the wall-fluid boundaries. In vitro investigations on two carotid bifurcation phantoms, normal and diseased, were conducted, and their relative differences in terms of the flow patterns and WSR distribution were demonstrated. It is shown that high-frame-rate UIV technique can be a non-invasive tool to measure quantitatively the spatio-temporal velocity and WSS distribution.

Vaporising phase change ultrasound contrast agent in microvascular confinement

The sub-micron perfluorocarbon-based phase change contrast agents (PCCAs) have shown great potential for imaging both microvasculature and tumour microenvironment. However the effect of geometrical confinement on the acoustic vaporisation has not been explored. We report the investigation of the difference in PCCA-produced contrast increase after activation with and without a microvessel confinement using in vitro experiment phantom. The results demonstrated that the acoustic vaporisation of PCCAs in the 200-micron cellulose tube was significantly different from that in the open environment, with contrast enhancement suppressed by up to several fold.

Surface Charge Measurement of SonoVue, Definity and Optison: A Comparison of Laser Doppler Electrophoresis and Micro-Electrophoresis

Microbubble (MB) contrast-enhanced ultrasonography is a promising tool for targeted molecular imaging. It is important to determine the MB surface charge accurately as it affects the MB interactions with cell membranes. In this article, we report the surface charge measurement of SonoVue, Definity and Optison. We compare the performance of the widely used laser Doppler electrophoresis with an in-house micro-electrophoresis system. By optically tracking MB electrophoretic velocity in a microchannel, we determined the zeta potentials of MB samples. Using micro-electrophoresis, we obtained zeta potential values for SonoVue, Definity and Optison of -28.3, -4.2 and -9.5 mV, with relative standard deviations of 5%, 48% and 8%, respectively. In comparison, laser Doppler electrophoresis gave -8.7, +0.7 and +15.8 mV with relative standard deviations of 330%, 29,000% and 130%, respectively. We found that the reliability of laser Doppler electrophoresis is compromised by MB buoyancy. Micro-electrophoresis determined zeta potential values with a 10-fold improvement in relative standard deviation.

Imaging of vaporised sub-micron phase change contrast agents with high frame rate ultrasound and optics

Phase-change ultrasound contrast agent (PCCA), or nanodroplet, shows promise as an alternative to the conventional microbubble agent over a wide range of diagnostic applications. Meanwhile, high-frame-rate (HFR) ultrasound imaging with microbubbles enables unprecedented temporal resolution compared to traditional contrast-enhanced ultrasound imaging. The combination of HFR ultrasound imaging and PCCAs can offer the opportunity to observe and better understand PCCA behaviour after vaporisation captures the fast phenomenon at a high temporal resolution. In this study, we utilised HFR ultrasound at frame rates in the kilohertz range (5-20 kHz) to image native and size-selected PCCA populations immediately after vaporisation in vitro within clinical acoustic parameters. The size-selected PCCAs through filtration are shown to preserve a sub-micron-sized (mean diameter  <  200 nm) population without micron-sized outliers (>1 µm) that originate from native PCCA emulsion. The results demonstrate imaging signals with different amplitudes and temporal features compared to that of microbubbles. Compared with the microbubbles, both the B-mode and pulse-inversion (PI) signals from the vaporised PCCA populations were reduced significantly in the first tens of milliseconds, while only the B-mode signals from the PCCAs were recovered during the next 400 ms, suggesting significant changes to the size distribution of the PCCAs after vaporisation. It is also shown that such recovery in signal over time is not evident when using size-selective PCCAs. Furthermore, it was found that signals from the vaporised PCCA populations are affected by the amplitude and frame rate of the HFR ultrasound imaging. Using high-speed optical camera observation (30 kHz), we observed a change in particle size in the vaporised PCCA populations exposed to the HFR ultrasound imaging pulses. These findings can further the understanding of PCCA behaviour under HFR ultrasound imaging.

Acoustic wave sparsely-activated localization microscopy (AWSALM): super-resolution ultrasound imaging using acoustic activation and deactivation of nanodroplets

Photo-activated localization microscopy (PALM) has revolutionized the field of fluorescence microscopy by breaking the diffraction limit in spatial resolution. In this study, “acoustic wave sparsely activated localization microscopy (AWSALM),” an acoustic counterpart of PALM, is developed to super-resolve structures which cannot be resolved by conventional B-mode imaging. AWSALM utilizes acoustic waves to sparsely and stochastically activate decafluorobutane nanodroplets by acoustic vaporization and to simultaneously deactivate the existing vaporized nanodroplets via acoustic destruction. In this method, activation, imaging, and deactivation are all performed using acoustic waves. Experimental results show that sub-wavelength micro-structures not resolvable by standard B-mode ultrasound images can be separated by AWSALM. This technique is flow independent and does not require a low concentration of contrast agents, as is required by current ultrasound super resolution techniques. Acoustic activation and deactivation can be controlled by adjusting the acoustic pressure, which remains well within the FDA approved safety range. In conclusion, this study shows the promise of a flow and contrast agent concentration independent super-resolution ultrasound technique which has potential to be faster and go beyond vascular imaging.

Dual frequency transcranial ultrasound for contrast enhanced ultrafast brain functional imaging

Ultrafast, high sensitivity Doppler imaging has led to the development of functional ultrasound (fUS), a novel functional imaging modality for detecting haemodynamic changes in cerebral blood vessels at high spatialtemporal resolution (∼100 μm, ∼1 ms). To allow for this blood tissue signal to be detected the skull either needs to be partially removed or thinned. To overcome this attenuation it has been shown that blood tissue signal can be enhanced using microbubble contrast agent, allowing for transcranial functional imaging to take place. By continuing to use a high frequency transmission, however, the attenuation in penetration will remain high and will limit imaging depths. Furthermore, by driving this microbubble oscillation with a frequency far greater than the resonance of the majority of microbubbles a suboptimal response is produced. We instead propose a dual frequency system which first transmits with a low frequency transducer to significantly decrease attenuation in transmit and to produce a stronger microbubble response, and to then receive with a second, higher frequency transducer set at an angle to detect the superharmonic component of the resulting broadband microbubble signal. From an ex vivo study it was found that the microbubble response for the dual frequency system increased relative to the conventional single frequency system with peak negative pressure (PNP) (starting from 140 kPa onwards) and showed excellent tissue suppression with a +12 dB gain in CTR when comparing the best performance of the two systems. This was followed by a promising in vivo pilot study demonstrating that the our dual frequency system was able to image cerebral blood flow in mice with intact skulls.

High frame rate ultrasound imaging of vaporised phase change contrast agents

Phase-change ultrasound contrast agents (PCCAs), have shown promises for ultrasound-mediated diagnostics and therapeutics. High-frame-rate ultrasound imaging with microbubbles significantly improved temporal resolution and contrast compared to conventional contrast-enhanced ultrasound imaging, offering opportunities to observe and better understand PCCA behavior after vaporisation. In this study, high-frame-rate ultrasound was used to image the PCCAs immediately after vaporisation in vitro with clinical ultrasound parameters. Compared with microbubbles, the vaporised PCCAs have shown different temporal signal features with high-frame-rate ultrasound imaging: a significant decrease in the first tens of milliseconds after acoustic vaporisation while no evident change over time with microbubble signal. Simultaneous optical microscopy was operated to observe behaviors of the vaporised PCCAs. Highframe-rate ultrasound imaging enables improved understanding of the acoustic behavior of the vaporised PCCA populations at high temporary resolution.