Ultrafast ultrasound localization microscopy for deep super-resolution vascular imaging.

Non-invasive imaging deep into organs at microscopic scales remains an open quest in biomedical imaging. Although optical microscopy is still limited to surface imaging owing to optical wave diffusion and fast decorrelation in tissue, revolutionary approaches such as fluorescence photo-activated localization microscopy led to a striking increase in resolution by more than an order of magnitude in the last decade. In contrast with optics, ultrasonic waves propagate deep into organs without losing their coherence and are much less affected by in vivo decorrelation processes. However, their resolution is impeded by the fundamental limits of diffraction, which impose a long-standing trade-off between resolution and penetration. This limits clinical and preclinical ultrasound imaging to a sub-millimetre scale. Here we demonstrate in vivo that ultrasound imaging at ultrafast frame rates (more than 500 frames per second) provides an analogue to optical localization microscopy by capturing the transient signal decorrelation of contrast agents--inert gas microbubbles. Ultrafast ultrasound localization microscopy allowed both non-invasive sub-wavelength structural imaging and haemodynamic quantification of rodent cerebral microvessels (less than ten micrometres in diameter) more than ten millimetres below the tissue surface, leading to transcranial whole-brain imaging within short acquisition times (tens of seconds). After intravenous injection, single echoes from individual microbubbles were detected through ultrafast imaging. Their localization, not limited by diffraction, was accumulated over 75,000 images, yielding 1,000,000 events per coronal plane and statistically independent pixels of ten micrometres in size. Precise temporal tracking of microbubble positions allowed us to extract accurately in-plane velocities of the blood flow with a large dynamic range (from one millimetre per second to several centimetres per second). These results pave the way for deep non-invasive microscopy in animals and humans using ultrasound. We anticipate that ultrafast ultrasound localization microscopy may become an invaluable tool for the fundamental understanding and diagnostics of various disease processes that modify the microvascular blood flow, such as cancer, stroke and arteriosclerosis.

Contrast enhanced ultrasound by real-time spatiotemporal filtering of ultrafast images.

Contrast enhanced ultrasound (CEUS) takes advantage of the nonlinear behaviour of injected microbubbles. If these contrast techniques yield good specificity between bubbles and tissues, they suffer some drawbacks, inherently linked to their dependence on nonlinear content. In recent years, plane-wave ultrasound reached frame rates of up to 20 000 fps. In this study we propose a linear technique for CEUS that takes advantage of these very high frame rates to separate bubbles from tissue without requiring nonlinearities. Data-driven spatiotemporal filtering operations are used to separate different features in the image on the basis of coherence both in space and time. Such filter recently proved to improve Doppler sensitivity (Demene et al 2015 IEEE Trans. Med. Imaging 34 2271-85). In contrast with bubbles, even slow moving ones, tissues are highly coherent both in space and time. Therefore, singular value decomposition (SVD) seems to be a powerful tool for the separation of contrast agents and tissues. In this paper, we apply SVD processing to linear ultrafast ultrasound images for CEUS Doppler. The contrast levels reached by this technique were compared to those of a nonlinear gold standard sequence (PMPI Doppler) through a flow phantom study. The SVD technique reached contrast-to-tissue ratios (CTR) up to 10 dB higher in vitro, and proved to be robust in terms of probe motion and slow flow. A trial was also conducted on a transplanted human kidney, already imaged by means of power Doppler (Claudon et al 1999 Am. J. Roentgenol. 173 41-6) and microbubbles (Kay et al 2009 Clin. Radiol. 64 1081-7). Contrast levels yielded by the SVD technique measured up to 13 dB higher than those of PMPI Doppler.

Resolution limits of ultrafast ultrasound localization microscopy.

As in other imaging methods based on waves, the resolution of ultrasound imaging is limited by the wavelength. However, the diffraction-limit can be overcome by super-localizing single events from isolated sources. In recent years, we developed plane-wave ultrasound allowing frame rates up to 20,000 fps. Ultrafast processes such as rapid movement or disruption of ultrasound contrast agents (UCA) can thus be monitored, providing us with distinct punctual sources that could be localized beyond the diffraction limit. We previously showed experimentally that resolutions beyond λ/10 can be reached in ultrafast ultrasound localization microscopy (uULM) using a 128 transducer matrix in reception. Higher resolutions are theoretically achievable and the aim of this study is to predict the maximum resolution in uULM with respect to acquisition parameters (frequency, transducer geometry, sampling electronics). The accuracy of uULM is the error on the localization of a bubble, considered a point-source in a homogeneous medium. The proposed model consists in two steps: determining the timing accuracy of the microbubble echo in radiofrequency data, then transferring this time accuracy into spatial accuracy. The simplified model predicts a maximum resolution of 40 µm for a 1.75 MHz transducer matrix composed of two rows of 64 elements. Experimental confirmation of the model was performed by flowing microbubbles within a 60 µm microfluidic channel and localizing their blinking under ultrafast imaging (500 Hz frame rate). The experimental resolution, determined as the standard deviation in the positioning of the microbubbles, was predicted within 6 µm (13%) of the theoretical values and followed the analytical relationship with respect to the number of elements and depth. Understanding the underlying physical principles determining the resolution of superlocalization will allow the optimization of the imaging setup for each organ. Ultimately, accuracies better than the size of capillaries are achievable at several centimeter depths.