3D Super-Resolution US Imaging of Rabbit Lymph Node Vasculature in Vivo by Using Microbubbles.

Background Variations in lymph node (LN) microcirculation can be indicative of metastasis. The identification and quantification of metastatic LNs remains essential for prognosis and treatment planning, but a reliable noninvasive imaging technique is lacking. Three-dimensional super-resolution (SR) US has shown potential to noninvasively visualize microvascular networks in vivo. Purpose To study the feasibility of three-dimensional SR US imaging of rabbit LN microvascular structure and blood flow by using microbubbles. Materials and Methods In vivo studies were carried out to image popliteal LNs of two healthy male New Zealand white rabbits aged 6-8 weeks. Three-dimensional, high-frame-rate, contrast material-enhanced US was achieved by mechanically scanning with a linear imaging probe. Individual microbubbles were identified, localized, and tracked to form three-dimensional SR images and super-resolved velocity maps. Acoustic subaperture processing was used to improve image contrast and to generate enhanced power Doppler and color Doppler images. Vessel size and blood flow velocity distributions were evaluated and assessed by using Student paired t test. Results SR images revealed microvessels in the rabbit LN, with branches clearly resolved when separated by 30 µm, which is less than half of the acoustic wavelength and not resolvable by using power or color Doppler. The apparent size distribution of most vessels in the SR images was below 80 µm and agrees with micro-CT data, whereas most of those detected with Doppler techniques were larger than 80 µm in the images. The blood flow velocity distribution indicated that most of the blood flow in rabbit popliteal LN was at velocities lower than 5 mm/sec. Conclusion Three-dimensional super-resolution US imaging using microbubbles allows noninvasive nonionizing visualization and quantification of lymph node microvascular structures and blood flow dynamics with resolution below the wave diffraction limit. This technology has potential for studying the physiologic functions of the lymph system and for clinical detection of lymph node metastasis. Published under a CC BY 4.0 license. Online supplemental material is available for this article.

Power Doppler Quantification in Assessing Gestational Trophoblastic Neoplasia.

PURPOSE: The FIGO score cannot accurately stratify low-risk gestational trophoblastic neoplasia (GTN) patients who develop chemoresistance to single agent methotrexate chemotherapy. Tumour vascularisation is a key risk factor and its quantification may provide non-invasive way of complementing risk assessment. MATERIALS AND METHODS: 187 FIGO-staged, low-risk GTN patients were prospectively recruited. Power Doppler ultrasound was analysed using a quantification program. Four diagnostic indicators were obtained comprising the number of colour pixels (NCP), mean dB, power Doppler quantification (PDQ), and percentage of colour pixels (%CP). Each indicator performance was assessed to determine if they could distinguish the subset of low-risk patients who became chemoresistant. RESULTS: There were 111 non-resistant and 76 resistant patients. NCP performed best at distinguishing these two groups where the non-resistant group had an average 3435 (±â€Š2060) pixels and the resistant group 6151 (±â€Š3192) pixels (p < 0.001). PDQ and %CP showed significant differences (p < 0.001) but had poorer performance (area under ROC curves were 72 % and 67 % respectively compared with 75 % for NCP). The mean dB index was not significantly different (p = 0.133). CONCLUSION: Power Doppler ultrasound quantification shows potential for non-invasive assessment of tumour vascularity and can distinguish low-risk GTN patients who become chemoresistant from those who have an uncomplicated course with first line treatment.

Mapping microbubble viscosity using fluorescence lifetime imaging of molecular rotors.

Encapsulated microbubbles are well established as highly effective contrast agents for ultrasound imaging. There remain, however, some significant challenges to fully realize the potential of microbubbles in advanced applications such as perfusion mapping, targeted drug delivery, and gene therapy. A key requirement is accurate characterization of the viscoelastic surface properties of the microbubbles, but methods for independent, nondestructive quantification and mapping of these properties are currently lacking. We present here a strategy for performing these measurements that uses a small fluorophore termed a "molecular rotor" embedded in the microbubble surface, whose fluorescence lifetime is directly related to the viscosity of its surroundings. We apply fluorescence lifetime imaging to show that shell viscosities vary widely across the population of the microbubbles and are influenced by the shell composition and the manufacturing process. We also demonstrate that heterogeneous viscosity distributions exist within individual microbubble shells even with a single surfactant component.

Tracking shear waves in turbid medium by light: theory, simulation, and experiment.

Shear wave propagation provides rich information for material mechanical characterization, including elasticity and viscosity. This Letter reports tracking of shear wave propagation in turbid media by laser-speckle-contrast analysis. The theory is described, and a Monte Carlo simulation of light shear wave interaction was developed. Simulation and experiments on tissue-mimicking phantoms agree well and show tracking of shear wave at the phantom surface and at depth as well as multiple shear waves interacting within the object. The relationship between speckle contrast value and shear wave amplitude is also investigated.

Prospects for enhancement of targeted radionuclide therapy of cancer using ultrasound.

Ultrasound-mediated drug delivery is a promising means of enhancing delivery, distribution and effectiveness of drugs within tumours. In this review, prospects for exploiting ultrasound to improve the tumour delivery and distribution of radiolabelled antibodies for radioimmunotherapy and to overcome barriers imposed by tumour microenvironment are discussed.

Ultrasonographic measures of synovitis in an early phase clinical trial: a double-blind, randomised, placebo and comparator controlled phase IIa trial of GW274150 (a selective inducible nitric oxide synthase inhibitor) in rheumatoid arthritis.

OBJECTIVES: To test the sensitivity to change of ultrasonographic endpoints in early phase clinical trials in subjects with active rheumatoid arthritis (RA). METHODS: A double-blind, placebo and comparator controlled, randomised, two-centre study investigated the effect on synovial thickness and vascularity of 28 days repeat daily oral dosing of 60 mg of the inducible nitric oxide synthase inhibitor GW274150 or 7.5 mg prednisolone in RA. Fifty patients with DAS28 scores ≥4.0 were assigned to 3 treatment arms of 17, 19 and 14 (on placebo, GW274150 and prednisolone respectively). Synovial thickness and vascularity of all 10 metacarpophalangeal joints were assessed by ultrasonography using a semi-quantitative scale at baseline (Day 1), Day 15 and Day 28. Vascularity was also measured quantitatively by power Doppler area. RESULTS: At Day 28, the GW274150 group showed a trend towards reduction in synovial thickness compared with placebo, with an adjusted mean decrease of 33% (p=0.072); the prednisolone group decreased significantly by 44% (p=0.011). Similarly, there was a trend to reduced synovial vascularity with GW274150 by 42% compared with placebo (p=0.075); prednisolone resulted in a statistically significant decrease of 55% (p=0.012). There was a 55% decrease in power Doppler area for GW274150, compared with placebo although the result was not statistically significant (p=0.375). Prednisolone 7.5 mg resulted in a highly statistically significant decrease of 95% (p=0.003). CONCLUSIONS: This study advocates the use of ultrasonographic measures of metacarpophalangeal joint synovitis as an endpoint for clinical studies assessing therapeutic potential of new compounds in small patient cohorts over 28 days.

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.

Effect of bubble shell nonlinearity on ultrasound nonlinear propagation through microbubble populations.

Nonlinear propagation of ultrasound through microbubble populations can generate artifacts and reduce contrast to tissue ratio in ultrasound imaging. The existing propagation model, which underestimates harmonic generation by an order of magnitude, was revised by incorporating a nonlinear constitutive equation for the coating into the description of the microbubble dynamics. Significantly better agreement with experiments was obtained, indicating that coating nonlinearity represents an important contribution to nonlinear propagation of ultrasound in microbubble populations. The results were found to be sensitive to the parameters characterizing the coating nonlinearity and thus accurate measurement of these parameters is required for accurate quantitative predictions.

Ring Artifact Correction for Phase-Insensitive Ultrasound Computed Tomography.

An algorithm was developed for the correction of ring artifacts in phase-insensitive ultrasound computed tomography attenuation images. Differences in the measurement sensitivity between the ultrasound transducer array elements cause discontinuities in the sinogram which manifest as rings and arcs in the reconstructed image. The magnitudes of the discontinuities are potentially time-varying and dependent on the attenuation being measured. The algorithm dynamically determines the measurement sensitivity of each transducer in the array during the scan by comparison with both the elements to its left and the elements to its right. Elements at either end of the array are corrected, assuming a zero-attenuation path. The two estimates of sensitivity are combined using a weighted mean similar to a Kalman filter. The algorithm was tested on simulated and experimentally acquired data. It was demonstrated to reduce the root-mean-square error (RMSE) of simulated images against ground-truth images by up to a factor of 50 compared with uncorrected images and to visibly reduce artifacts on images reconstructed from the experimentally acquired data.

Impact of Aperture, Depth, and Acoustic Clutter on the Performance of Coherent Multi-Transducer Ultrasound Imaging.

Transducers with a larger aperture size are desirable in ultrasound imaging to improve resolution and image quality. A coherent multi-transducer ultrasound imaging system (CoMTUS) enables an extended effective aperture through the coherent combination of multiple transducers. In this study, the discontinuous extended aperture created by CoMTUS and its performance for deep imaging and through layered media are investigated by both simulations and experiments. Typical image quality metrics-resolution, contrast and contrast-to-noise ratio-are evaluated and compared with a standard single probe imaging system. Results suggest that the image performance of CoMTUS depends on the relative spatial location of the arrays. The resulting effective aperture significantly improves resolution, while the separation between the arrays may degrade contrast. For a limited gap in the effective aperture (less than a few centimetres), CoMTUS provides benefits to image quality compared to the standard single probe imaging system. Overall, CoMTUS shows higher sensitivity and reduced loss of resolution with imaging depth. In general, CoMTUS imaging performance was unaffected when imaging through a layered medium with different speed of sound values and resolution improved up to 80% at large imaging depths.

High-Frame-Rate Tri-Plane Echocardiography With Spiral Arrays: From Simulation to Real-Time Implementation.

Major cardiovascular diseases (CVDs) are associated with (regional) dysfunction of the left ventricle. Despite the 3-D nature of the heart and its dynamics, the assessment of myocardial function is still largely based on 2-D ultrasound imaging, thereby making diagnosis heavily susceptible to the operator's expertise. Unfortunately, to date, 3-D echocardiography cannot provide adequate spatiotemporal resolution in real-time. Hence, tri-plane imaging has been introduced as a compromise between 2-D and true volumetric ultrasound imaging. However, tri-plane imaging typically requires high-end ultrasound systems equipped with fully populated matrix array probes embedded with expensive and little flexible electronics for two-stage beamforming. This article presents an advanced ultrasound system for real-time, high frame rate (HFR), and tri-plane echocardiography based on low element count sparse arrays, i.e., the so-called spiral arrays. The system was simulated, experimentally validated, and implemented for real-time operation on the ULA-OP 256 system. Five different array configurations were tested together with four different scan sequences, including multi-line and planar diverging wave transmission. In particular, the former can be exploited to achieve, in tri-plane imaging, the same temporal resolution currently used in clinical 2-D echocardiography, at the expenses of contrast (-3.5 dB) and signal-to-noise ratio (SNR) (-8.7 dB). On the other hand, the transmission of planar diverging waves boosts the frame rate up to 250 Hz, but further compromises contrast (-10.5 dB), SNR (-9.7 dB), and lateral resolution (+46%). In conclusion, despite an unavoidable loss in image quality and sensitivity due to the limited number of elements, HFR tri-plane imaging with spiral arrays is shown to be feasible in real-time and may enable real-time functional analysis of all left ventricular segments of the heart.

3-D Super-Resolution Ultrasound Imaging With a 2-D Sparse Array.

High-frame-rate 3-D ultrasound imaging technology combined with super-resolution processing method can visualize 3-D microvascular structures by overcoming the diffraction-limited resolution in every spatial direction. However, 3-D super-resolution ultrasound imaging using a full 2-D array requires a system with a large number of independent channels, the design of which might be impractical due to the high cost, complexity, and volume of data produced. In this study, a 2-D sparse array was designed and fabricated with 512 elements chosen from a density-tapered 2-D spiral layout. High-frame-rate volumetric imaging was performed using two synchronized ULA-OP 256 research scanners. Volumetric images were constructed by coherently compounding nine-angle plane waves acquired at a pulse repetition frequency of 4500 Hz. Localization-based 3-D super-resolution images of two touching subwavelength tubes were generated from 6000 volumes acquired in 12 s. Finally, this work demonstrates the feasibility of 3-D super-resolution imaging and super-resolved velocity mapping using a customized 2-D sparse array transducer.

Optimal Control of SonoVue Microbubbles to Estimate Hydrostatic Pressure.

The measurement of cardiac and aortic pressures enables diagnostic insight into cardiac contractility and stiffness. However, these pressures are currently assessed invasively using pressure catheters. It may be possible to estimate these pressures less invasively by applying microbubble ultrasound contrast agents as pressure sensors. The aim of this study was to investigate the subharmonic response of the microbubble ultrasound contrast agent SonoVue (Bracco Spa, Milan, Italy) at physiological pressures using a static pressure phantom. A commercially available cell culture cassette with Luer connections was used as a static pressure chamber. SonoVue was added to the phantom, and radio frequency data were recorded on the ULtrasound Advanced Open Platform (ULA-OP). The mean subharmonic amplitude over a 40% bandwidth was extracted at 0-200-mmHg hydrostatic pressures, across 1.7-7.0-MHz transmit frequencies and 3.5%-100% maximum scanner acoustic output. The Rayleigh-Plesset equation for single-bubble oscillations and additional hysteresis experiments were used to provide insight into the mechanisms underlying the subharmonic pressure response of SonoVue. The subharmonic amplitude of SonoVue increased with hydrostatic pressure up to 50 mmHg across all transmit frequencies and decreased thereafter. A decreasing microbubble surface tension may drive the initial increase in the subharmonic amplitude of SonoVue with hydrostatic pressure, while shell buckling and microbubble destruction may contribute to the subsequent decrease above 125-mmHg pressure. In conclusion, a practical operating regime that may be applied to estimate cardiac and aortic blood pressures from the subharmonic signal of SonoVue has been identified.

Poisson Statistical Model of Ultrasound Super-Resolution Imaging Acquisition Time.

A number of acoustic super-resolution techniques have recently been developed to visualize microvascular structure and flow beyond the diffraction limit. A crucial aspect of all ultrasound (US) super-resolution (SR) methods using single microbubble localization is time-efficient detection of individual bubble signals. Due to the need for bubbles to circulate through the vasculature during acquisition, slow flows associated with the microcirculation limit the minimum acquisition time needed to obtain adequate spatial information. Here, a model is developed to investigate the combined effects of imaging parameters, bubble signal density, and vascular flow on SR image acquisition time. We find that the estimated minimum time needed for SR increases for slower blood velocities and greater resolution improvement. To improve SR from a resolution of λ /10 to λ /20 while imaging the microvasculature structure modeled here, the estimated minimum acquisition time increases by a factor of 14. The maximum useful imaging frame rate to provide new spatial information in each image is set by the bubble velocity at low blood flows (<150 mm/s for a depth of 5 cm) and by the acoustic wave velocity at higher bubble velocities. Furthermore, the image acquisition procedure, transmit frequency, localization precision, and desired super-resolved image contrast together determine the optimal acquisition time achievable for fixed flow velocity. Exploring the effects of both system parameters and details of the target vasculature can allow a better choice of acquisition settings and provide improved understanding of the completeness of SR information.

Investigation of Microbubble Detection Methods for Super-Resolution Imaging of Microvasculature.

Ultrasound super-resolution techniques use the response of microbubble (MB) contrast agents to visualize the microvasculature. Techniques that localize isolated bubble signals first require detection algorithms to separate the MB and tissue responses. This work explores the three main MB detection techniques for super-resolution of microvasculature. Pulse inversion (PI), differential imaging (DI), and singular value decomposition (SVD) filtering were compared in terms of the localization accuracy, precision, and contrast-to-tissue ratio. MB responses were simulated based on the properties of Sonovue and using the Marmottant model. Nonlinear propagation through tissue was modeled using the k-Wave software package. For the parameters studied, the results show that PI is most appropriate for low frequency applications, but also most dependent on transducer bandwidth. SVD is preferable for high frequency acquisition where localization precision on the order of a few microns is possible. PI is largely independent of flow direction and speed compared to SVD and DI, so is appropriate for visualizing the slowest flows and tortuous vasculature. SVD is unsuitable for stationary MBs and can introduce a localization error on the order of hundreds of microns over the speed range 0-2 mm/s and flow directions from lateral (parallel to probe) to axial (perpendicular to probe). DI is only suitable for flow rates >0.5 mm/s or as flow becomes more axial. Overall, this study develops an MB and tissue nonlinear simulation platform to improve understanding of how different MB detection techniques can impact the super-resolution process and explores some of the factors influencing the suitability of each.

Coherent Multi-Transducer Ultrasound Imaging.

This work extends the effective aperture size by coherently compounding the received radio frequency data from multiple transducers. As a result, it is possible to obtain an improved image, with enhanced resolution, an extended field of view (FoV), and high-acquisition frame rates. A framework is developed in which an ultrasound imaging system consisting of N synchronized matrix arrays, each with partly shared FoV, take turns to transmit plane waves (PWs). Only one individual transducer transmits at each time while all N transducers simultaneously receive. The subwavelength localization accuracy required to combine information from multiple transducers is achieved without the use of any external tracking device. The method developed in this study is based on the study of the backscattered echoes received by the same transducer and resulting from a targeted scatterer point in the medium insonated by the multiple ultrasound probes of the system. The current transducer locations along with the speed of sound in the medium are deduced by optimizing the cross correlation between these echoes. The method is demonstrated experimentally in 2-D for two linear arrays using point targets and anechoic lesion phantoms. The first demonstration of a free-hand experiment is also shown. Results demonstrate that the coherent multi-transducer ultrasound imaging method has the potential to improve ultrasound image quality, improving resolution, and target detectability. Compared with coherent PW compounding using a single probe, lateral resolution improved from 1.56 to 0.71 mm in the coherent multi-transducer imaging method without acquisition frame rate sacrifice (acquisition frame rate 5350 Hz).

Fast Acoustic Wave Sparsely Activated Localization Microscopy (fast-AWSALM): Ultrasound Super-Resolution using Plane-Wave Activation of Nanodroplets.

Localization-based ultrasound super-resolution imaging using microbubble contrast agents and phase-change nano-droplets has been developed to visualize microvascular structures beyond the diffraction limit. However, the long data acquisition time makes the clinical translation more challenging. In this study, fast acoustic wave sparsely activated localization microscopy (fast-AWSALM) was developed to achieve super-resolved frames with sub-second temporal resolution, by using low-boiling-point octafluoropropane nanodroplets and high frame rate plane waves for activation, destruction, as well as imaging. Fast-AWSALM was demonstrated on an in vitro microvascular phantom to super-resolve structures that could not be resolved by conventional B-mode imaging. The effects of the temperature and mechanical index on fast-AWSALM was investigated. Experimental results show that sub-wavelength micro-structures as small as 190 lm were resolvable in 200 ms with plane-wave transmission at a center frequency of 3.5 MHz and a pulse repetition frequency of 5000 Hz. This is about a 3.5 fold reduction in point spread function full-width-half-maximum compared to that measured in conventional B-mode, and two orders of magnitude faster than the recently reported AWSALM under a non-flow/very slow flow situations and other localization based methods. Just as in AWSALM, fast-AWSALM does not require flow, as is required by current microbubble based ultrasound super resolution techniques. In conclusion, this study shows the promise of fast-AWSALM, a super-resolution ultrasound technique using nanodroplets, which can generate super-resolution images in milli-seconds and does not require flow.

3-D Motion Correction for Volumetric Super-Resolution Ultrasound Imaging.

Motion during image acquisition can cause image degradation in all medical imaging modalities. This is particularly relevant in 2-D ultrasound imaging, since out-of-plane motion can only be compensated for movements smaller than elevational beamwidth of the transducer. Localization based super-resolution imaging creates even a more challenging motion correction task due to the requirement of a high number of acquisitions to form a single super-resolved frame. In this study, an extension of two-stage motion correction method is proposed for 3-D motion correction. Motion estimation was performed on high volumetric rate ultrasound acquisitions with a handheld probe. The capability of the proposed method was demonstrated with a 3-D microvascular flow simulation to compensate for handheld probe motion. Results showed that two-stage motion correction method reduced the average localization error from 136 to 18 µm.

Two-Stage Motion Correction for Super-Resolution Ultrasound Imaging in Human Lower Limb.

The structure of microvasculature cannot be resolved using conventional ultrasound (US) imaging due to the fundamental diffraction limit at clinical US frequencies. It is possible to overcome this resolution limitation by localizing individual microbubbles through multiple frames and forming a superresolved image, which usually requires seconds to minutes of acquisition. Over this time interval, motion is inevitable and tissue movement is typically a combination of large- and small-scale tissue translation and deformation. Therefore, super-resolution (SR) imaging is prone to motion artifacts as other imaging modalities based on multiple acquisitions are. This paper investigates the feasibility of a two-stage motion estimation method, which is a combination of affine and nonrigid estimation, for SR US imaging. First, the motion correction accuracy of the proposed method is evaluated using simulations with increasing complexity of motion. A mean absolute error of 12.2 was achieved in simulations for the worst-case scenario. The motion correction algorithm was then applied to a clinical data set to demonstrate its potential to enable in vivo SR US imaging in the presence of patient motion. The size of the identified microvessels from the clinical SR images was measured to assess the feasibility of the two-stage motion correction method, which reduced the width of the motion-blurred microvessels to approximately 1.5-fold.

High-Frame-Rate Contrast Echocardiography Using Diverging Waves: Initial In Vitro and In Vivo Evaluation.

Contrast echocardiography (CE) ultrasound with microbubble contrast agents has significantly advanced our capability for assessment of cardiac function, including myocardium perfusion quantification. However, in standard CE techniques obtained with line by line scanning, the frame rate and image quality are limited. Recent research has shown significant frame-rate improvement in noncontrast cardiac imaging. In this work, we present and initially evaluate, both in vitro and in vivo, a high-frame-rate (HFR) CE imaging system using diverging waves and pulse inversion sequence. An imaging frame rate of 5500 frames/s before and 250 frames/s after compounding is achieved. A destruction-replenishment sequence has also been developed. The developed HFR CE is compared with standard CE in vitro on a phantom and then in vivo on a sheep heart. The image signal-to-noise ratio and contrast between the myocardium and the chamber are evaluated. The results show up to 13.4-dB improvement in contrast for HFR CE over standard CE when compared at the same display frame rate even when the average spatial acoustic pressure in HFR CE is 36% lower than the standard CE. It is also found that when coherent compounding is used, the HFR CE image intensity can be significantly modulated by the flow motion in the chamber.