Efficient Image Registration for Ultrasound Localization Microscopy by Obtaining Gradients via Integration Across Iterations

Tissue motion correction through image registration is essential for ultrasound localization microscopy (ULM). Parametric image registration is commonly formulated as an optimization problem where motion parameters are iteratively updated to maximize image similarity, and used optimization algorithms typically rely on gradient information, the explicit evaluation of which can become computationally demanding. This work investigates Extremum Seeking Control (ESC) as an alternative to explicit derivative evaluation in image registration. By obtaining descent information via integrating perturbed and demodulated image similarity metric across iterations, ESC avoids differentiation of the image similarity metric with respect to motion parameters in each iteration. The classical ESC, whose optimization behavior approximates that of classical gradient descent (GD), is first compared with GD for affine image registration using simulated ground-truth motions derived from a beating ex vivo porcine heart dataset. The results show that ESC achieves registration accuracy and convergence behavior comparable to GD while reducing per-iteration computational cost by approximately 3.5-fold. ESC is subsequently employed in a two-stage motion correction pipeline, where affine registration compensates for global tissue motion and B-spline registration corrects residual local deformation. The proposed method is applied to ULM imaging of a beating ex vivo porcine heart and achieves a spatial resolution of 219 um, substantially below the half-wavelength diffraction limit of 321 um associated with 2.4 MHz diverging-wave imaging. These results demonstrate that ESC provides an effective alternative to explicit derivative evaluation in ULM image registration, enabling accurate motion correction and high-quality super-resolution imaging.

Reconstructing 3D Neural Hemodynamics using Sparse Ultrasound Localization Microscopy Data

Ultrasound Localization Microscopy (ULM) has presented great potential in functional imaging, benefiting from its ability to reconstruct deep microvasculature. However, the hemodynamic reconstruction is compromised by sparsity in the ULM data, as a limited number of MB tracks cannot sample the complete speed profile in one vessel. Here, we propose to reconstruct hemodynamics using sparse ULM velocity maps by solving a laminar flow model through stochastic variational inference. In addition to vascular geometry and flow velocity maps, the proposed method generates two new ULM maps - a pressure gradient map and a map describing uncertainty of the estimation. By investigating the effect of sparsity in ULM maps on the quantification and visualization of hemodynamics, we demonstrate the effectiveness of the proposed method in dealing with sparse ULM maps via simulations and 3D rat brain imaging. Accurately reconstructing a broad range of hemodynamic parameters and associate uncertanties using sparse ULM data may help detect subtle and dynamic brain activity.

CycleULM: A unified label-free deep learning framework for ultrasound localisation microscopy

Super-resolution ultrasound via microbubble (MB) localisation and tracking, also known as ultrasound localisation microscopy (ULM), can resolve microvasculature beyond the acoustic diffraction limit. However, significant challenges remain in localisation performance and data acquisition and processing time. Deep learning methods for ULM have shown promise to address these challenges, however, they remain limited by in vivo label scarcity and the simulation-to-reality domain gap. We present CycleULM, the first unified label-free deep learning framework for ULM. CycleULM learns a physics-emulating translation between the real contrast-enhanced ultrasound (CEUS) data domain and a simplified MB-only domain, leveraging the power of CycleGAN without requiring paired ground truth data. With this translation, CycleULM removes dependence on high-fidelity simulators or labelled data, and makes MB localisation and tracking substantially easier. Deployed as modular plug-and-play components within existing pipelines or as an end-to-end processing framework, CycleULM delivers substantial performance gains across both in silico and in vivo datasets. Specifically, CycleULM improves image contrast (contrast-to-noise ratio) by up to 15.3 dB and sharpens CEUS resolution with a 2.5{\times} reduction in the full width at half maximum of the point spread function. CycleULM also improves MB localisation performance, with up to +40% recall, +46% precision, and a -14.0 μm mean localisation error, yielding more faithful vascular reconstructions. Importantly, CycleULM achieves real-time processing throughput at 18.3 frames per second with order-of-magnitude speed-ups (up to ~14.5{\times}). By combining label-free learning, performance enhancement, and computational efficiency, CycleULM provides a practical pathway toward robust, real-time ULM and accelerates its translation to clinical applications.

Determining the utility of ultrafast nonlinear contrast enhanced and super resolution ultrasound for imaging microcirculation in the human small intestine

The regulation of intestinal blood flow is critical to gastrointestinal function. Imaging the intestinal mucosal micro-circulation in vivo has the potential to provide new insight into the gut physiology and pathophysiology. We aimed to determine whether ultrafast contrast enhanced ultrasound (CEUS) and super-resolution ultrasound localisation microscopy (SRUS/ULM) could be a useful tool for imaging the small intestine microcirculation in vivo non-invasively and for detecting changes in blood flow in the duodenum. Ultrafast CEUS and SRUS/ULM were used to image the small intestinal microcirculation in a cohort of 20 healthy volunteers (BMI<25). Participants were imaged while conscious and either having been fasted, or following ingestion of a liquid meal or water control, or under acute stress. For the first time we have performed ultrafast CEUS and ULM on the human small intestine, providing unprecedented resolution images of the intestinal microcirculation. We evaluated flow speed inside small vessels in healthy volunteers (2.78 +/- 0.05 mm/s, mean +/- SEM) and quantified changes in the perfusion of this microcirculation in response to nutrient ingestion. Perfusion of the microvasculature of the intestinal mucosa significantly increased post-prandially (36.2% +/- 12.2%, mean +/- SEM, p<0.05). The feasibility of 3D SRUS/ULM was also demonstrated. This study demonstrates the potential utility of ultrafast CEUS for assessing perfusion and detecting changes in blood flow in the duodenum. SRUS/ULM also proved a useful tool to image the microvascular blood flow in vivo non-invasively and to evaluate blood speed inside the microvasculature of the human small intestine.

Online 4D Ultrasound-Guided Robotic Tracking Enables 3D Ultrasound Localisation Microscopy with Large Tissue Displacements

Super-Resolution Ultrasound (SRUS) imaging through localising and tracking microbubbles, also known as Ultrasound Localisation Microscopy (ULM), has demonstrated significant potential for reconstructing microvasculature and flows with sub-diffraction resolution in clinical diagnostics. However, imaging organs with large tissue movements, such as those caused by respiration, presents substantial challenges. Existing methods often require breath holding to maintain accumulation accuracy, which limits data acquisition time and ULM image saturation. To improve image quality in the presence of large tissue movements, this study introduces an approach integrating high-frame-rate ultrasound with online precise robotic probe control. Tested on a microvasculature phantom with translation motions up to 20 mm, twice the aperture size of the matrix array used, our method achieved real-time tracking of the moving phantom and imaging volume rate at 85 Hz, keeping majority of the target volume in the imaging field of view. ULM images of the moving cross channels in the phantom were successfully reconstructed in post-processing, demonstrating the feasibility of super-resolution imaging under large tissue motions. This represents a significant step towards ULM imaging of organs with large motion.

Enhancing super-resolution ultrasound localisation through multi-frame deconvolution exploiting spatiotemporal coherence

Super-resolution ultrasound imaging through microbubble (MB) localisation and tracking, also known as ultrasound localisation microscopy, allows non-invasive sub-diffraction resolution imaging of microvasculature in animals and humans. The number of MBs localised from the acquired contrast-enhanced ultrasound (CEUS) images and the localisation precision directly influence the quality of the resulting super-resolution microvasculature images. However, non-negligible noise present in the CEUS images can make localising MBs challenging. To enhance the MB localisation performance, we propose a Multi-Frame Deconvolution (MF-Decon) framework that can exploit the spatiotemporal coherence inherent in the CEUS data, with new spatial and temporal regularisers designed based on total variation (TV) and regularisation by denoising (RED). Based on the MF-Decon framework, we introduce two novel methods: MF-Decon with spatial and temporal TVs (MF-Decon+3DTV) and MF-Decon with spatial RED and temporal TV (MF-Decon+RED+TV). Results from in silico simulations indicate that our methods outperform two widely used methods using deconvolution or normalised cross-correlation across all evaluation metrics, including precision, recall, $F_1$ score, mean and standard localisation errors. In particular, our methods improve MB localisation precision by up to 39% and recall by up to 12%. Super-resolution microvasculature maps generated with our methods on a publicly available in vivo rat brain dataset show less noise, better contrast, higher resolution and more vessel structures.

Ultrafast 3-D Super Resolution Ultrasound using Row-Column Array specific Coherence-based Beamforming and Rolling Acoustic Sub-aperture Processing: In Vitro, In Vivo and Clinical Study

The row-column addressed array is an emerging probe for ultrafast 3-D ultrasound imaging. It achieves this with far fewer independent electronic channels and a wider field of view than traditional 2-D matrix arrays, of the same channel count, making it a good candidate for clinical translation. However, the image quality of row-column arrays is generally poor, particularly when investigating tissue. Ultrasound localisation microscopy allows for the production of super-resolution images even when the initial image resolution is not high. Unfortunately, the row-column probe can suffer from imaging artefacts that can degrade the quality of super-resolution images as `secondary' lobes from bright microbubbles can be mistaken as microbubble events, particularly when operated using plane wave imaging. These false events move through the image in a physiologically realistic way so can be challenging to remove via tracking, leading to the production of 'false vessels'. Here, a new type of rolling window image reconstruction procedure was developed, which integrated a row-column array-specific coherence-based beamforming technique with acoustic sub-aperture processing for the purposes of reducing `secondary' lobe artefacts, noise and increasing the effective frame rate. Using an {\it{in vitro}} cross tube, it was found that the procedure reduced the percentage of `false' locations from $\sim$26\% to $\sim$15\% compared to traditional orthogonal plane wave compounding. Additionally, it was found that the noise could be reduced by $\sim$7 dB and that the effective frame rate could be increased to over 4000 fps. Subsequently, {\it{in vivo}} ultrasound localisation microscopy was used to produce images non-invasively of a rabbit kidney and a human thyroid.

Acceleration-Based Kalman Tracking for Super-Resolution Ultrasound Imaging in vivo

Super-resolution ultrasound can image microvascular structure and flow at sub-wave-diffraction resolution based on localising and tracking microbubbles. Currently, tracking microbubbles accurately under limited imaging frame rates and high microbubble concentrations remains a challenge, especially under the effect of cardiac pulsatility and in highly curved vessels. In this study, an acceleration-incorporated microbubble motion model is introduced into a Kalman tracking framework. The tracking performance was evaluated using simulated microvasculature with different microbubble motion parameters and acquisition frame rates, and in vivo human breast tumour ultrasound datasets. The simulation results show that the acceleration-based method outperformed the non-acceleration-based method at different levels of acceleration and acquisition frame rates and achieved significant improvement in true positive rate (up to 10.03%), false negative rate (up to 28.61%) and correctly pairing fraction (up to 170.14%). The proposed method can also reduce errors in vasculature reconstruction via the acceleration-based nonlinear interpolation, compared with linear interpolation (up to 19 um). The tracking results from temporally downsampled low frame rate in vivo datasets from human breast tumours show that the proposed method has better microbubble tracking performance than the baseline method, if using results from the initial high frame data as reference. Finally, the acceleration estimated from tracking results also provides a spatial speed gradient map that may contain extra valuable diagnostic information.

Transthoracic super-resolution ultrasound localisation microscopy of myocardial vasculature in patients

Micro-vascular flow in the myocardium is of significant importance clinically but remains poorly understood. Up to 25% of patients with symptoms of coronary heart diseases have no obstructive coronary arteries and have suspected microvascular diseases. However, such microvasculature is difficult to image in vivo with existing modalities due to the lack of resolution and sensitivity. Here, we demonstrate the feasibility of transthoracic super-resolution ultrasound localisation microscopy (SRUS/ULM) of myocardial microvasculature and hemodynamics in a large animal model and in patients, using a cardiac phased array probe with a customised data acquisition and processing pipeline. A multi-level motion correction strategy was proposed. A tracking framework incorporating multiple features and automatic parameter initialisations was developed to reconstruct microcirculation. In two patients with impaired myocardial function, we have generated SRUS images of myocardial vascular structure and flow with a resolution that is beyond the wave-diffraction limit (half a wavelength), using data acquired within a breath hold. Myocardial SRUS/ULM has potential to improve the understanding of myocardial microcirculation and the management of patients with cardiac microvascular diseases.

3D Super-Resolution Ultrasound with Adaptive Weight-Based Beamforming

Super-resolution ultrasound (SRUS) imaging through localising and tracking sparse microbubbles has been shown to reveal microvascular structure and flow beyond the wave diffraction limit. Most SRUS studies use standard delay and sum (DAS) beamforming, where large main lobe and significant side lobes make separation and localisation of densely distributed bubbles challenging, particularly in 3D due to the typically small aperture of matrix array probes. This study aims to improve 3D SRUS by implementing a low-cost 3D coherence beamformer based on channel signal variance, as well as two other adaptive weight-based coherence beamformers: nonlinear beamforming with p-th root compression and coherence factor. The 3D coherence beamformers, together with DAS, are compared in computer simulation, on a microflow phantom, and in vivo. Simulation results demonstrate that the adaptive weight-based beamformers can significantly narrow the main lobe and suppress the side lobes for modest computational cost. Significantly improved 3D SR images of microflow phantom and a rabbit kidney are obtained through the adaptive weight-based beamformers. The proposed variance-based beamformer performs best in simulations and experiments.