Search

Functional ultrasound (fUS) is a neuroimaging method that uses ultrasound to track changes in cerebral blood volume as an indirect readout of neuronal activity at high spatiotemporal resolution. fUS is capable of imaging head-fixed or freely behaving rodents and of producing volumetric images of the entire mouse brain. It has been applied to many species, including primates and humans. Now that fUS is reaching maturity, it is being adopted by the neuroscience community. However, the nature of the fUS signal and the different implementations of fUS are not necessarily accessible to nonspecialists. This review aims to introduce these ultrasound concepts to all neuroscientists. We explain the physical basis of the fUS signal and the principles of the method, present the state of the art of its hardware implementation, and give concrete examples of current applications in neuroscience. Finally, we suggest areas for improvement during the next few years.

The brain is composed of a dense and ramified vascular network comprising various sizes of arteries, veins, and capillaries. One way to assess the risk of cerebrovascular pathologies is to use computational models to predict the physiological effects of a reduction of blood supply and correlate these responses with observations of brain damage. Therefore, it is crucial to establish a detailed 3D organization of the brain vasculature, which could be used to develop more accurate in silico models. For this purpose, we have adapted our open-access functional ultrasound imaging platform previously designed for recording brain-wide activity that is now capable of fast and reproducible acquisition, segmentation, and reconstruction of the cortical vasculature. For the first time, it allows us to digitize the cortical vasculature in awake rodents with a ∼100 µm3 spatial resolution. Contrary to most available strategies, our approach can be performed in vivo within minutes. Moreover, it is easy to implement since it neither requires exogenous contrast agents nor long post-processing time. Hence, we performed a cortex-wide reconstruction of the vasculature and its quantitative analysis, including i) classification of descending arteries versus ascending veins in more than 1500 vessels/animal, ii) quick estimation of their length. Importantly, we confirmed the relevance of our approach in a model of cortical stroke, which enables quick visualization of the ischemic lesion. This development contributes to extending the capabilities of ultrasound neuroimaging to understand better cerebrovascular pathologies such as stroke, vascular cognitive impairment, and brain tumors and is highly scalable for the clinic.

Ischemic stroke occurs with no warning, and therefore, very little is known about hemodynamic perturbations in the brain immediately after stroke onset. Here, functional ultrasound imaging was used to monitor variations in relative cerebral blood volume (rCBV) compared to baseline. rCBV levels were analyzed brain-wide and continuously at high spatiotemporal resolution (100μm, 2Hz) until 70mins after stroke onset in rats. We compared two stroke models, with either a permanent occlusion of the middle cerebral artery (MCAo) or a tandem occlusion of both the common carotid and middle cerebral arteries (CCAo+MCAo). We observed a typical hemodynamic pattern, including a quick drop of the rCBV after MCAo, followed by spontaneous reperfusion of several brain regions located in the vicinity of the ischemic core. The severity and location of the ischemia were highly variable between animals. Still, both parameters were, on average, in good agreement with the final ischemic lesion volume measured 24hrs after stroke onset for the MCAo but not the CCAo+MCAo model. For the latter, we observed that the infarct was extended to regions that were initially not ischemic and located rostrally of the ischemic core. These regions strongly colocalize with the origin of transient hemodynamic events associated with spreading depolarizations.

During social interactions, individuals influence each other to coordinate their actions. Vocal communication is an exceptionally efficient way to exert such influence. Where and how social interactions are dynamically modulated in the brain is unknown. We used functional ultrasound imaging in marmoset monkeys – a highly vocal species - to investigate the dynamics of medial social brain areas in vocal perception, production, and audio-vocal interaction. We found that the activity of a distributed network of subcortical and cortical regions distinguishes calls associated with different social contexts. This same brain network showed different dynamics during externally and internally driven vocalizations. These findings suggest the existence of a social-vocal brain network in medial cortical and subcortical areas that is fundamental in social communication.One Sentence SummaryA network of medial subcortical and cortical brain areas gate social communication in primates.

Most brain functions engage a network of distributed regions. Full investigation of these functions thus requires assessment of whole brains; however, whole-brain functional imaging of behaving animals remains challenging. This protocol describes how to follow brain-wide activity in awake head-fixed mice using functional ultrasound imaging, a method that tracks cerebral blood volume dynamics. We describe how to set up a functional ultrasound imaging system with a provided acquisition software (miniScan), establish a chronic cranial window (timing surgery: ~3–4 h) and image brain-wide activity associated with a stimulus at high resolution (100 × 110 × 300 µm and 10 Hz per brain slice, which takes ~45 min per imaging session). We include codes that enable data to be registered to a reference atlas, production of 3D activity maps, extraction of the activity traces of ~250 brain regions and, finally, combination of data from multiple sessions (timing analysis averages ~2 h). This protocol enables neuroscientists to observe global brain processes in mice. Cerebral blood volume dynamics in awake head-fixed mice are tracked via functional ultrasound imaging through a chronic cranial window, giving a high-resolution measure of brain-wide activity.

Functional ultrasound imaging (fUSI) is an appealing method for measuring blood flow and thus infer brain activity, but it relies on the physiology of neurovascular coupling and requires extensive signal processing. To establish to what degree fUSI trial-by-trial signals reflect neural activity, we performed simultaneous fUSI and neural recordings with Neuropixels probes in awake mice. fUSI signals strongly correlated with the slow (

Imaging large-scale circuit dynamics is crucial to understanding brain function, but most techniques have a limited depth of field. Here, we describe volumetric functional ultrasound imaging (vfUSI), a platform for brain-wide vfUSI of hemodynamic activity in awake head-fixed mice. We combined a high-frequency 1,024-channel 2D-array transducer with advanced multiplexing and high-performance computing for real-time 3D power Doppler imaging at a high spatiotemporal resolution (220 × 280 × 175 μm3, up to 6 Hz). We developed a standardized software pipeline for registration, segmentation, and temporal analysis in 268 individual brain regions based on the Allen Mouse Common Coordinate Framework. We demonstrated the high sensitivity of vfUSI under multiple experimental conditions, and we successfully imaged stimulus-evoked activity when only a few trials were averaged. We also mapped neural circuits in vivo across the whole brain during optogenetic activation of specific cell types. Moreover, we identified the sequential activation of sensory-motor networks during a grasping water-droplet task. ispartof: NEURON vol:108 issue:5 pages:861-+ ispartof: location:United States status: published

Neuronal cell-types are arranged in brain-wide circuits to guide behavior. In mice, the superior colliculus is comprised of a set of cell-types that each innervate distinct downstream targets. Here we reveal the brain-wide networks downstream of four collicular cell-types by combining functional ultrasound imaging (fUSi) with optogenetics to monitor neural activity at a resolution of ~100 μm. Each neuronal group triggered different behaviors, and activated distinct, partially overlapping sets of brain nuclei. This included regions not previously thought to mediate defensive behaviors, e.g. the posterior paralaminar nuclei of the thalamus (PPnT), that we show to play a role in suppressing habituation. Electrophysiological recordings support the fUSi findings and show that neurons in the downstream nuclei preferentially respond to innately threatening visual stimuli. This work provides insight into the functional organization of the networks governing defensive behaviors and demonstrates an experimental approach to explore the whole-brain neuronal activity downstream of targeted cell-types.

SUMMARYImaging of large-scale circuit dynamics is crucial to gain a better understanding of brain function, but most techniques have a limited depth of field. Here we describe vfUSI, a platform for brain-wide volumetric functional ultrasound imaging of hemodynamic activity in awake head-fixed mice. We combined high-frequency 1024-channel 2D-array transducer with advanced multiplexing and high-performance computing for real-time 3D Power Doppler imaging at high spatiotemporal resolution (220×280×175-μm3 voxel size, up to 6 Hz). In addition, we developed a standardized software pipeline for registration and segmentation based on the Allen Mouse Common Coordinate Framework, allowing for temporal analysis in 268 individual brain regions. We demonstrate the high sensitivity of vfUSI in multiple experimental situations where stimulus-evoked activity can be recorded using a minimal number of trials. We also mapped neural circuits in vivo across the whole brain during optogenetic activation of specific cell-types. Moreover, we revealed the sequential activation of sensory-motor regions during a grasping water droplet task. vfUSI will become a key neuroimaging technology because it combines ease of use, reliability, and affordability.

Developing efficient brain imaging technologies by combining a high spatiotemporal resolution and a large penetration depth is a key step for better understanding the neurovascular interface that emerges as a main pathway to neurodegeneration in many pathologies such as dementia. This review focuses on the advances in two complementary techniques: multi-photon laser scanning microscopy (MPLSM) and functional ultrasound imaging (fUSi). MPLSM has become the gold standard for in vivo imaging of cellular dynamics and morphology, together with cerebral blood flow. fUSi is an innovative imaging modality based on Doppler ultrasound, capable of recording vascular brain activity over large scales (i.e., tens of cubic millimeters) at unprecedented spatial and temporal resolution for such volumes (up to 10 μm pixel size at 10 kHz). By merging these two technologies, researchers may have access to a more detailed view of the various processes taking place at the neurovascular interface. MPLSM and fUSi are also good candidates for addressing the major challenge of real-time delivery, monitoring, and in vivo evaluation of drugs in neuronal tissue

Following middle cerebral artery (MCA) stroke, enhanced contralesional evoked responses have been consistently reported both in man and rodents as part of plastic processes thought to influence motor recovery. How early this marker of large-scale network reorganization develops has however been little addressed, yet has clinical relevance for rehabilitation strategies targeting plasticity. Previous work in mice has reported enhanced contralesional responses to unaffected-side forepaw stimulation as early as 45 min after MCA small branch occlusion. Using functional ultrasound imaging (fUSi) in anesthetized rats subjected to distal temporary MCA occlusion (MCAo), we assessed here (i) whether enhanced contralesional responses also occurred with unaffected-side whisker pad stimulation, and if so, how early after MCAo; and (ii) the time course of this abnormal response during occlusion and after reperfusion. We replicate in a more proximal MCA occlusion model the earlier findings of ultra-early enhanced contralesional evoked responses. In addition, we document this phenomenon within minutes after MCAo, and its persistence throughout the entire 90-min occlusion as well as 90-min reperfusion periods studied. These findings suggest that plastic processes may start within minutes following MCAo in rodents. If replicated in man, they might have implications regarding how early plasticity-enhancing therapies can be initiated after stroke. ispartof: JOURNAL OF CEREBRAL BLOOD FLOW AND METABOLISM vol:38 issue:10 pages:1690-1700 ispartof: location:United States status: published

Innovative imaging methods help to investigate the complex relationship between brain activity and behavior in freely moving animals. Functional ultrasound (fUS) is an imaging modality suitable for recording cerebral blood volume (CBV) dynamics in the whole brain but has so far been used only in head-fixed and anesthetized rodents. We designed a fUS device for tethered brain imaging in freely moving rats based on a miniaturized ultrasound probe and a custom-made ultrasound scanner. We monitored CBV changes in rats during various behavioral states such as quiet rest, after whisker or visual stimulations, and in a food-reinforced operant task. We show that fUS imaging in freely moving rats could efficiently decode brain activity in real time.

Functional ultrasound imaging is a method recently developed to assess brain activity via hemodynamics in rodents. Doppler ultrasound signals allow the measurement of cerebral blood volume (CBV) and red blood cells' (RBCs') velocity in small vessels. However, this technique originally requires performing a large craniotomy that limits its use to acute experiments only. Moreover, a detailed description of the hemodynamic changes that underlie functional ultrasound imaging has not been described but is essential for a better interpretation of neuroimaging data.To overcome the limitation of the craniotomy, we developed a dedicated thinned skull surgery for chronic imaging. This procedure did not induce brain inflammation nor neuronal death as confirmed by immunostaining. We successfully acquired both high-resolution images of the microvasculature and functional movies of the brain hemodynamics on the same animal at 0, 2, and 7days without loss of quality. Then, we investigated the spatiotemporal evolution of the CBV hemodynamic response function (HRF) in response to sensory-evoked electrical stimulus (1mA) ranging from 1 (200μs) to 25 pulses (5s). Our results indicate that CBV HRF parameters such as the peak amplitude, the time to peak, the full width at half-maximum and the spatial extent of the activated area increase with stimulus duration. Functional ultrasound imaging was sensitive enough to detect hemodynamic responses evoked by only a single pulse stimulus. We also observed that the RBC velocity during activation could be separated in two distinct speed ranges with the fastest velocities located in the upper part of the cortex and slower velocities in deeper layers. For the first time, functional ultrasound imaging demonstrates its potential to image brain activity chronically in small animals and offers new insights into the spatiotemporal evolution of cerebral hemodynamics.

Although conventional pulse-wave Doppler has proved to be a valuable diagnostic method for many vascular pathologies, it is hampered by issues related to repeatability as well as problems associated with quantification and system-dependent variability. These limitations are due to intrinsic spectral broadening on the Doppler spectrum, resulting from the directivity pattern of the ultrasound focused beam. Here, we develop a new spatial statistical technique, Doppler frequency spatial analysis (DFSA), which is based on ultrafast plane-wave imaging. Similar to standard pulse-wave Doppler, which is commonly used by sonographers, it yields a two-dimensional output (frequency versus time), while dramatically reducing the presence of intrinsic spectral broadening on the Doppler spectra. Therefore, the technique is much more sensitive to the velocity profile and turbulences than the standard pulse-wave Doppler. The proposed technique could improve diagnosis of vascular diseases, including arterial plaque characterization. Moreover, by summarizing all main information contained in the ultrafast Doppler acquisition, it permits a direct visualization of the data within the velocity profile. Here, we have compared our novel statistical technique to the standard pulse-wave Doppler approach during in vivo imaging of the human carotid artery. Notably, we achieved a greater than 4-fold reduction in intrinsic spectral broadening.

Topographic representation of the outside world is a key feature of sensory systems, but so far it has been difficult to define how the activity pattern of the olfactory information is distributed at successive stages in the olfactory system. We studied odor-evoked activation patterns in the main olfactory bulb and the anterior piriform cortex of rats using functional ultrasound (fUS) imaging. fUS imaging is based on the use of ultrafast ultrasound scanners and detects variations in the local blood volume during brain activation. It makes deep brain imaging of ventral structures, such as the piriform cortex, possible. Stimulation with two different odors (hexanal and pentylacetate) induced the activation of odor-specific zones that were spatially segregated in the main olfactory bulb. Interestingly, the same odorants triggered the activation of the entire anterior piriform cortex, in all layers, with no distinguishable odor-specific areas detected in the power Doppler images. These fUS imaging results confirm the spatial distribution of odor-evoked activity in the main olfactory bulb, and furthermore, they reveal the absence of such a distribution in the anterior piriform cortex at the macroscopic scale in vivo.

High-intensity focused ultrasound causes selective tissue necrosis efficiently and safely, namely, in the prostate, liver, and uterine fibroid. Nevertheless, ablation of brain tissue using focused ultrasound remains limited due to strong aberrations induced by the skull. To achieve ultrasonic transcranial brain ablation, such aberrations have to be compensated. In this study, non-invasive therapy was performed on monkeys using adaptive correction of the therapeutic beam and 3D simulations of transcranial wave propagation based on 3D computed tomographic (CT) scan information. The aim of the study was two-fold: induce lesions in a non-human primate brain non-invasively and investigate the potential side effects. Stereotactic targeting was performed on five Macaca fascicularis individuals. Each hemisphere was treated separately with a 15-day interval and animals were sacrificed two days after the last treatment. The ultrasonic dose delivered at the focus was increased from one treatment location to the other to estimate the thermal dose for tissue alteration. Thermal doses in the brain were determined by numerical computations. Treatment efficiency and safety were evaluated histologically. The threshold for tissue damage in the brain was measured to be between 90 and 280 cumulative equivalent minutes at 43 °C. Intravenous injection of corticoids before the treatment limited the side effects.

Introduction: Following MCAo, tissue outcome varies depending on depth and duration of hypoperfusion and efficiency of reperfusion. However, the precise time-course of these events in relation to tissue and behavioral outcome remains unsettled due to lack of a wide field-of-view quantitative imaging technique able to map perfusion in the rodent brain at high spatiotemporal resolution. Here we used fUS, a novel approach to map cerebral blood volume (CBV) without contrast agent. Hypothesis: fUS will allow quantitative, near real-time mapping of CVB during and after tMCAo. Methods: 45min filament tMCAo was induced in adult SD rats; sham rats were also used. fUS was used to map the penetrating arterioles and venules of the ipsi- and contra-lateral motor (M1-2) and somatosensory (S1) cortex in coronal sections across the MCA territory at 80μm resolution. Three-min coronal scans were taken at different levels before, during and immediately after MCAo, and at 3 and 6 days thanks to a thinned-skull preparation. CBV was expressed relative to mirror ROI. In addition, a 1-hr movie (one frame/5s) was taken starting a few mins after reperfusion. Serial Neuroscore and 2 sensorimotor tasks were given over 3w post-MCAo, and then NeuN, IBa1 and GFAP immunofluorescence (IF) at post-mortem. Results: fUS showed a ∼80% CBV reduction in S1 during occlusion (p Conclusions: fUS efficiently mapped the acute changes in CBV during occlusion and following reperfusion with high spatio-temporal resolution, allowing the charting of fine tissue reperfusion dynamics in the individual rat. fUS is ideal to longitudinally map real-time cerebral perfusion in experimental stroke from the hyper-acute through to the chronic stage.

A combination of radiation force and ultrafast ultra-sound imaging is used to both generate and track the propagation of a shear wave in the brain whose local speed is directly related to stiffness, characterized by the dynamic shear modulus G*. When performed on trepanated rats, this approach called shear wave imaging (SWI) provides 3-D brain elasticity maps reaching a spatial resolution of 0.7 mm × 1 mm × 0.4 mm with a good reproducibility (

Doppler-based flow analysis methods require acquisition of ultrasound data at high spatio-temporal sampling rates. These rates represent a major technical challenge for ultrasound systems because a compromise between spatial and temporal resolution must be made in conventional approaches. Consequently, ultrasound scanners can either provide full quantitative Doppler information on a limited sample volume (spectral Doppler), or averaged Doppler velocity and/or power estimation on a large region of interest (Doppler flow imaging). In this work, we investigate a different strategy for acquiring Doppler information that can overcome the limitations of the existing Doppler modes by significantly reducing the required acquisition time. This technique is called ultrafast compound Doppler imaging and is based on the following concept: instead of successively insonifying the medium with focused beams, several tilted plane waves are sent into the medium and the backscattered signals are coherently summed to produce high-resolution ultrasound images. We demonstrate that this strategy allows reduction of the acquisition time by a factor of up to of 16 while keeping the same Doppler performance. Depending on the application, different directions to increase performance of Doppler analysis are proposed and the improvement is quantified: the ultrafast compound Doppler method allows faster acquisition frame rates for high-velocity flow imaging, or very high sensitivity for low-flow applications. Full quantitative Doppler flow analysis can be performed on a large region of interest, leading to much more information and improved functionality for the physician. By leveraging the recent emergence of ultrafast parallel beamforming systems, this paper demonstrates that breakthrough performances in flow analysis can be reached using this concept of ultrafast compound Doppler.

Summary Large numbers of brain regions are active during behaviors. A high-resolution, brain-wide activity map could identify brain regions involved in specific behaviors. We have developed functional ultrasound imaging to record whole-brain activity in behaving mice at a resolution of ∼100 μm. We detected 87 active brain regions during visual stimulation that evoked the optokinetic reflex, a visuomotor behavior that stabilizes the gaze both horizontally and vertically. Using a genetic mouse model of congenital nystagmus incapable of generating the horizontal reflex, we identified a subset of regions whose activity was reflex dependent. By blocking eye motion in control animals, we further separated regions whose activity depended on the reflex’s motor output. Remarkably, all reflex-dependent but eye motion-independent regions were located in the thalamus. Our work identifies functional modules of brain regions involved in sensorimotor integration and provides an experimental approach to monitor whole-brain activity of mice in normal and disease states., Highlights • Functional ultrasound enables imaging whole-brain activity during mouse behavior • Activity in 87 brain regions are modulated during the optokinetic reflex • Reflex-related regions were identified by perturbing retinal direction selectivity • A subset of these regions, all in the thalamus, are independent of eye motion, Macé et al. developed an approach for imaging whole-brain activity during mouse behavior using ultrasound. They revealed many brain regions activated during the optokinetic reflex, a visuomotor behavior that stabilizes gaze. They parsed them into functional modules using reflex perturbations.

The emergence of ultrafast frame rates in ultrasonic imaging has been recently made possible by the development of new imaging modalities such as transient elastography. Data acquisition rates reaching more than thousands of images per second enable the real-time visualization of shear mechanical waves propagating in biological tissues, which convey information about local viscoelastic properties of tissues. The first proposed approach for reaching such ultrafast frame rates consists of transmitting plane waves into the medium. However, because the beamforming process is then restricted to the receive mode, the echographic images obtained in the ultrafast mode suffer from a low quality in terms of resolution and contrast and affect the robustness of the transient elastography mode. It is here proposed to improve the beamforming process by using a coherent recombination of compounded plane-wave transmissions to recover high-quality echographic images without degrading the high frame rate capabilities. A theoretical model is derived for the comparison between the proposed method and the conventional B-mode imaging in terms of contrast, signal-to-noise ratio, and resolution. Our model predicts that a significantly smaller number of insonifications, 10 times lower, is sufficient to reach an image quality comparable to conventional B-mode. Theoretical predictions are confirmed by in vitro experiments performed in tissue-mimicking phantoms. Such results raise the appeal of coherent compounds for use with standard imaging modes such as B-mode or color flow. Moreover, in the context of transient elastography, ultrafast frame rates can be preserved while increasing the image quality compared with flat insonifications. Improvements on the transient elastography mode are presented and discussed.

In vivo assessment of dispersion affecting the propagation of visco-elastic waves in soft tissues is key to understand the rheology of human tissues. In this paper, the ability of the supersonic shear imaging (SSI) technique to generate planar shear waves propagating in tissues is fully exploited. First, by strongly limiting shear wave diffraction in the imaging plane, this imaging technique enables to discriminate between the usually concomitant influences of both medium rheological properties and diffraction affecting the shear wave dispersion. Second, transient propagation of these plane shear waves in soft tissues can be measured using echographic images acquired at very high frame. In vitro and in vivo experiments demonstrate that dispersion curves, which characterize the rheological behavior of tissues by measuring the frequency dependence of shear wave speed and attenuation, can be recovered in the 75-600 Hz frequency range. Based on a phase difference algorithm, the dispersion curves are computed in 1 cm2 regions of interest from the acquired propagation movie. In vivo measurements in Biceps Brachii muscle and liver of three healthy volunteers show important differences in the rheological behavior of these different tissues. Liver tissue appears to be much more dispersive with a phase velocity ranging from approximately 1.5 m/s at 75 Hz to approximately 3 m/s at 500 Hz whereas muscle tissue shows an important anisotropy, shear waves propagating longitudinally to the muscular fibers are almost nondispersive while those propagating transversally are very dispersive with a shear wave speed ranging from 0.5 to 2 m/s between 75 and 500 Hz. The estimation of dispersion curves is local and can be performed separately in different regions of the organ. This signal processing approach based on the SSI modality introduces the new concept of in vivo shear wave spectroscopy (SWS) that could become an additional tool for tissue characterization. This paper demonstrates the in vivo ability of this SWS to quantify both local shear elasticity and dispersion in real time.

Pulse-inversion (PI) sequences are sensitive to the nonlinear echoes from microbubbles allowing an improvement in the blood-to-tissue contrast. However, at larger mechanical indices, this contrast is reduced by harmonics produced during nonlinear propagation. A method for tissue harmonics cancellation exploiting time reversal is experimentally implemented using a 128-channel 12-bit emitter receiver. The probe calibration is performed by acquiring the nonlinear echo of a wire in water. These distorted pulses are time-reversed, optimized and used for the PI imaging of a tissue phantom. Compared to normal (straight) pulses, the time-reversed distorted pulses reduced the tissue signal in PI by 11 dB. The second harmonic signals from microbubbles flowing in a wall-less vessel were unaffected by the correction. This technique can thus increase the blood-to-tissue contrast ratio while keeping the pressure and the number of pulses constant.

This paper presents an initial clinical evaluation of in vivo elastography for breast lesion imaging using the concept of supersonic shear imaging. This technique is based on the combination of a radiation force induced in tissue by an ultrasonic beam and an ultrafast imaging sequence capable of catching in real time the propagation of the resulting shear waves. The local shear wave velocity is recovered using a time-offlight technique and enables the 2-D mapping of shear elasticity. This imaging modality is implemented on a conventional linear probe driven by a dedicated ultrafast echographic device. Consequently, it can be performed during a standard echographic examination. The clinical investigation was performed on 15 patients, which corresponded to 15 lesions (4 cases BI-RADS 3, 7 cases BI-RADS 4 and 4 cases BI-RADS 5). The ability of the supersonic shear imaging technique to provide a quantitative and local estimation of the shear modulus of abnormalities with a millimetric resolution is illustrated on several malignant (invasive ductal and lobular carcinoma) and benign cases (fibrocystic changes and viscous cysts). In the investigated cases, malignant lesions were found to be significantly different from benign solid lesions with respect to their elasticity values. Cystic lesions have shown no shear wave propagate at all in the lesion (because shear waves do not propage in liquid). These preliminary clinical results directly demonstrate the clinical feasibility of this new elastography technique in providing quantitative assessment of relative stiffness of breast tissues. This technique of evaluating tissue elasticity gives valuable information that is complementary to the B-mode morphologic information. More extensive studies are necessary to validate the assumption that this new mode potentially helps the physician in both false-positive and false-negative rejection.

Bursts of focused ultrasound energy a billion times more intense than diagnostic ultrasound have become a non-invasive option for tumor ablation, from prostate cancer to uterine fibroid, during the last decade. Despite this progress, many issues still need to be addressed. First, for brain targeting, the correction of distortions induced by the skull remains today a technological achievement that still needs to be validated clinically. Secondly, the problem of motion artifacts for abdominal treatments becomes today an important research topic. For all these issues, the potential of new ultrasonic therapy devices able to work both in Transmit and Receive modes will be emphasized and clinical results on monkeys and pigs will be presented.

Elastic waves propagating inside a solid chaotic cavity create a diffusive random field that contains both longitudinal and shear waves. In the current paper, we are interested in the field radiated in a fluid in contact with such cavity. The goal of this paper is to predict the spatial focusing properties that can be obtained in the fluid using a time-reversal piezoelectric transducer in contact with the cavity. We present a statistical approach that supposes a fully diffused wavefield inside the cavity with an equipartition of energy between longitudinal and shear waves. We show that the critical angles of transmission in the solid–fluid interface generate a cut-off of the spatial frequencies and then a degradation in the spatial focusing. This limitation can be overcome using a rough surface. A set of experiments conducted in the MHz range confirm the theoretical model.

Bursts of focused ultrasound energy a thousand times more intense than diagnostic ultrasound have become a non-invasive option for treating cancer, from breast to prostate or uterine fibroid, during the last decade. Despite this progress, many issues still need to be addressed. First, the distortions caused by defocusing obstacles, such as the skull or ribs, on the ultrasonic therapeutic beam are still being investigated. Multi-element transducer technology must be used in order to achieve such transcranial or transcostal adaptive focusing. Second, the problem of motion artifacts, a key component in the treatment of abdominal lesions, has been shown significantly to influence the efficacy and treatment time. Though many methods have been proposed for the detection of organ motion, little work has been done to develop a comprehensive solution including motion tracking and feedback correction in real time. This paper is a review of the work achieved by authors in transcranial high-intensity focused ultrasound (HIFU), transcostal HIFU and motion compensated HIFU. For these three issues, the optimal solution can be reached using the same technology of multi-element transducers devices able to work both in transmit and receive modes.

Following middle cerebral artery occlusion, tissue outcome ranges from normal to infarcted depending on depth and duration of hypoperfusion as well as occurrence and efficiency of reperfusion. However, the precise time course of these changes in relation to tissue and behavioral outcome remains unsettled. To address these issues, a three-dimensional wide field-of-view and real-time quantitative functional imaging technique able to map perfusion in the rodent brain would be desirable. Here, we applied functional ultrasound imaging, a novel approach to map relative cerebral blood volume without contrast agent, in a rat model of brief proximal transient middle cerebral artery occlusion to assess perfusion in penetrating arterioles and venules acutely and over six days thanks to a thinned-skull preparation. Functional ultrasound imaging efficiently mapped the acute changes in relative cerebral blood volume during occlusion and following reperfusion with high spatial resolution (100 µm), notably documenting marked focal decreases during occlusion, and was able to chart the fine dynamics of tissue reperfusion (rate: one frame/5 s) in the individual rat. No behavioral and only mild post-mortem immunofluorescence changes were observed. Our study suggests functional ultrasound is a particularly well-adapted imaging technique to study cerebral perfusion in acute experimental stroke longitudinally from the hyper-acute up to the chronic stage in the same subject.

Retrieving the out-of-plane blood flow velocity vector from two-dimensional transverse acquisitions of large vessels could improve the quantification of flow rate and maximum speed. The in-plane vector flow component can be computed easily using the Doppler frequency shift. The main problem is estimating the angle between the probe imaging plane and the vessel axis to derive the out-of-plane component from in-plane measurements. In this article, we study the case in which the velocity vector can be decomposed on two directions: the out-of-plane direction and the in-plane depth direction. We explore the combination of a technique called intrinsic spectral broadening with ultrafast plane wave imaging to retrieve the out-of-plane component of the flow velocity vector. Using a one-time probe calibration of this intrinsic spectral broadening, out-of-plane angle and flow speed can be recovered easily, thus avoiding approximations of a complex theoretical analysis. For the calibration step, ultrafast plane wave imaging permits a fast calibration procedure for the Doppler intrinsic spectral broadening. In vitro experimental validations are performed on a homogeneous flow phantom and a Poiseuille flow; the absolute speed was retrieved with 6% error. The potential of the technique is demonstrated in vivo on the human carotid artery. Combined with in-plane vector flow approaches, this out-of-plane Doppler imaging method paves the way to threedimensional vector flow imaging using only conventional onedimensional probe technology.

The design of two-dimensional (2-D) arrays for three-dimensional (3-D) ultrasonic imaging is a major challenge in medical and nondestructive applications. Thousands of transducers are typically needed for focusing and steering in a 3-D volume. In this article, we propose a different concept allowing us to obtain electronic 3-D focusing with a small number of transducers. The basic idea is to couple a small number of transducers to a chaotic reverberating cavity with one face in contact with the body of the patient. The reverberations of the ultrasonic waves inside the cavity create at each reflection virtual transducers. The cavity acts as an ultrasonic kaleidoscope multiplying the small number of transducers and creating a much larger virtual transducer array. By exploiting time-reversal processing, it is possible to use collectively all the virtual transducers to focus a pulse everywhere in a 3-D volume. The reception process is based on a nonlinear pulse-inversion technique in order to ensure a good contrast. The feasibility of this concept for the building of 3-D images was demonstrated using a prototype relying only on 31 emission transducers and a single reception transducer.

In order to achieve an optimal focusing through heterogeneous media we need to build the inverse filter of the propagation operator. Time reversal is an easy and robust way to achieve such an inverse filter in nondissipative media. However, as soon as losses appear in the medium, time reversal is not equivalent to the inverse filter anymore. Consequently, it does not produce the optimal focusing and beam degradations may appear. In such cases, we showed in previous works that the optimal focusing can be recovered by using the so-called spatiotemporal inverse filter technique. This process requires the presence of a complete set of receivers inside the medium. It allows one to reach the optimal focusing even in extreme situations such as ultrasonic focusing through human skull or audible sound focusing in strongly reverberant rooms. But, this technique is time consuming and implied fastidious numerical calculations. In this paper we propose a new way to process this inverse filter focusing technique in real time and without any calculation. The new process is based on iterative time reversal process. Contrary to the classical inverse filter technique, this iteration does not require any computation and achieves the inverse filter in an experimental way using wave propagation instead of computational power. The convergence from time reversal to inverse filter during the iterative process is theoretically explained. Finally, the feasibility of this iterative technique is experimentally demonstrated for ultrasound applications.

The use of piezoelectric transducer arrays has opened up the possibility of electronic steering and focusing of acoustic beams to track kidney stones. However, owing to the limited pressure delivered by each transducer (typically 10 bar), the number of transducers needed to reach an amplitude at the focus on the order of 1000 bars is typically of some hundreds of elements. We present here a new solution based on 1-bit time reversal in a solid waveguide to obtain, with a small number of transducers, a very high amplitude pulse in tissues located in front of the waveguide. The idea is to take advantage of the temporal dispersion in the waveguide to create, after time reversal, a temporally recompressed pulse with a stronger amplitude. The aim of this work is threefold: first, we experimentally demonstrate 1-bit time reversal between a point source in water and several transducers fastened to one section of a finite-length cylindrical waveguide. Second, we numerically and experimentally study the temporal an...

Retrieving the blood flow velocity vector on 2D acquisitions transverse to the vessel could improve the quantification of flow rate in large vessels. The in plane component is easily computed using the Doppler frequency shift. The main problem is to retrieve the angle between the probe imaging plane and the vessel axis to compute the out of plane component. Here, we study the case where the speed vector can be decomposed on two directions: the out of plane direction and the depth direction. We explore the combination of a technique called spectral broadening with ultrafast plane wave imaging. Due to its high spatio temporal sampling, ultrafast plane wave imaging permits to quantify the Doppler spectrum of flows in each pixel. Moreover, ultrafast plane wave imaging permits a fast calibration method of Doppler spectra spectral broadening avoiding approximations of a complex theoretical analysis. An experimental demonstration of the method is performed in vivo on healthy human carotid artery.

Hemodynamic changes in the brain are often used as surrogates of neuronal activity to infer the loci of brain activity. A major limitation of conventional Doppler ultrasound for the imaging of these changes is that it is not sensitive enough to detect the blood flow in small vessels where the major part of the hemodynamic response occurs. Here, we present a μDoppler ultrasound method able to detect and map the cerebral blood volume (CBV) over the entire brain with an important increase in sensitivity. This method is based on imaging the brain at an ultrafast frame rate (1 kHz) using compounded plane wave emissions. A theoretical model demonstrates that the gain in sensitivity of the μDoppler method is due to the combination of 1) the high signal-to-noise ratio of the gray scale images, resulting from the synthetic compounding of backscattered echoes; and 2) the extensive signal averaging enabled by the high temporal sampling of ultrafast frame rates. This μDoppler imaging is performed in vivo on trepanned rats without the use of contrast agents. The resulting images reveal detailed maps of the rat brain vascularization with an acquisition time as short as 320 ms per slice. This new method is the basis for a real-time functional ultrasound (fUS) imaging of the brain.

A novel retrieval algorithm for the refractive-index profile of optical fibers from transverse interferograms is presented. This algorithm allows a quick and simple index-profile reconstruction even in the presence of measurement noise and near the fiber axis. The response to noise of different retrieval algorithms is discussed and computer simulations are presented.

Ultrafast plane wave imaging enables the acquisition of thousands of temporal samples per second over a large field of view compared to the scarce 100 frames per second of a conventional focused mode. However, a main problem remains: How to compute with this huge amount of data relevant information for diagnosis? In this article, we present a statistical technique that analyses the spatio-temporal distribution of the Doppler frequency. This statistical technique gives a two dimensional output frequency versus time as the standard pulse wave Doppler commonly used by sonographers but reduces dramatically the presence of geometric broadening on Doppler spectrums. Thereby, this technique becomes highly sensitive to the type of the flow profile and turbulences which could help to diagnosis vascular diseases as plaques in the carotid artery. By summarizing all the main information contained in the ultrafast acquisition it permits a direct visualization of the flow speed and its gradient. We compare this statistical technique and a standard pulse wave Doppler in vivo on the carotid artery.

Focusing a wave through heterogeneous media is an important problem in medical ultrasound imaging. In such aberrating media, in the presence of a small number of point reflectors, iterative time reversal is a well-known method able to focus on the strongest reflector. However, in presence of speckle noise generated by many non-resolved scatterers, iterative time reversal alone does not work. In this paper, we propose the use of the echoes coming from moving particles in a flow, such as red blood cells, to generate a virtual point reflector by iterative time reversal. The construction of the virtual point reflector is performed by a coherent addition of independent realizations of speckle coming from moving particles. After focusing on a virtual point reflector, ultrasound images can be locally corrected inside an isoplanatic patch. An application for the correction of power Doppler images is presented. A theoretical analysis shows that this iterative method allows focusing on the point of maximal insonification of the uncorrected beam.

Imaging intramyocardial vascular flows in real-time could strongly help to achieve better diagnostic of cardiovascular diseases. To date, no standard imaging modality allows describing accurately myocardial blood flow dynamics with good spatial and temporal resolution. We recently introduced a novel ultrasonic Doppler imaging technique based on compounded plane waves transmissions at ultrafast frame rate. The high sensitivity of this ultrafast Doppler technique permits to image the intramyocardial blood flow and its dynamics. A dedicated demodulation-filtering process is implemented to compensate for the large tissue velocity of the myocardium during the cardiac cycle. A signed power Doppler processing provides the discrimination between arterial and venous flows. Experiments were performed in vivo in a large animal open chest model ( N = 5 sheep) using a conventional ultrasonic probe placed at the surface of the heart. Results show the capability of the technique to image intramyocardial vascular flows in normal physiological conditions with good spatial (200 μm) and temporal resolution (10 ms). Flow dynamics over the cardiac cycle were investigated and the imaging method demonstrated a phase opposition of flow waveforms between arterial and venous flows. Finally, ultrafast Doppler combined with tissue motion compensation was found able to reveal vascular flow disruption in ischemic regions during occlusion of the main diagonal coronary artery.

Blood flow interruption in a cerebral artery causes brain ischemia and induces dramatic changes of perfusion and metabolism in the corresponding territory. We performed in parallel positron emission tomography (PET) with [15O]H2O, single photon emission computed tomography (SPECT) with [99mTc]hexamethylpropylene-amino-oxime ([99mTc]HMPAO) and ultrasonic ultrafast shear wave imaging (SWI) during, immediately after, and 1, 2, 4, and 7 days after middle cerebral artery occlusion (MCAO) in rats. Positron emission tomography and SPECT showed initial hypoperfusion followed by recovery at immediate reperfusion, hypoperfusion at day 1, and hyperperfusion at days 4 to 7. Hyperperfusion interested the whole brain, including nonischemic areas. Immunohistochemical analysis indicated active angiogenesis at days 2 to 7, strongly suggestive that hyperperfusion was supported by an increase in microvessel density in both brain hemispheres after ischemia. The SWI detected elastic changes of cerebral tissue in the ischemic area as early as day 1 after MCAO appearing as a softening of cerebral tissue whose local internal elasticity decreased continuously from day 1 to 7. Taken together, these results suggest that hyperperfusion after cerebral ischemia is due to formation of neovessels, and indicate that brain softening is an early and continuous process. The SWI is a promising novel imaging method for monitoring the evolution of cerebral ischemia over time in animals.

The building of high-power ultrasonic sources from piezoelectric ceramics is limited by the maximum voltage that the ceramics can endure. We have conceived a device that uses a small number of piezoelectric transducers fastened to a cylindrical metallic waveguide. A one-bit time- reversal operation transforms the long-lasting low-level dispersed wave forms into a sharp pulse, thus taking advantage of dispersion to generate high-power ultrasound. The pressure amplitude that is generated at the focus is found to be 15 times greater than that achieved with comparable standard techniques. Applications to lithotripsy are discussed and the destructive efficiency of the system is demonstrated on pieces of chalk.