In plane quantification ofin vivomuscle elastic anisotropy factor by steered ultrasound pushing beams.

Objective.Skeletal muscles are organized into distinct layers and exhibit anisotropic characteristics across various scales. Assessing the arrangement of skeletal muscles may provide valuable biomarkers for diagnosing muscle-related pathologies and evaluating the efficacy of clinical interventions.Approach. In this study, we propose a novel ultrafast ultrasound sequence constituted of steered pushing beams was proposed for ultrasound elastography applications in transverse isotropic muscle. Based on the propagation of the shear wave vertical mode, it is possible to fit the experimental results to retrieve in the same imaging plane, the shear modulus parallel to fibers as well as the elastic anisotropy factor (ratio of Young's moduli times the shear modulus perpendicular to fibers).Main results. The technique was demonstratedin vitroin phantoms andex vivoin fusiform beef muscles. At last, the technique was appliedin vivoon fusiform muscles (biceps brachii) and mono-pennate muscles (gastrocnemius medialis) during stretching and contraction.Significance. This novel sequence provides access to new structural and mechanical biomarkers of muscle tissue, including the elastic anisotropy factor, within the same imaging plane. Additionally, it enables the investigation of multiples parameters during muscle active and passive length changes.

Unravelling anisotropic nonlinear shear elasticity in muscles: Towards a non-invasive assessment of stress in living organisms.

Acoustoelasticity theory describes propagation of shear waves in uniaxially stressed medium and allows the retrieval of nonlinear elastic coefficients of tissues. In transverse isotropic medium such as muscles the theory leads to 9 different configurations of propagating shear waves (stress axis vs. fibers axis vs. shear wave polarization axis vs. shear wave propagation axis). In this work we propose to use 4 configurations to quantify these nonlinear parameters ex vivo and in vivo. Ex vivo experiments combining ultrasound shear wave elastography and mechanical testing were conducted on iliopsoas pig muscles to quantify three third-order nonlinear coefficients A, H and K that are possibly linked to the architectural structure of muscles. In vivo experiments were performed with human volunteers on biceps brachii during a stretching exercise on an ergometer. A combination of the third order nonlinear elastic parameters was assessed. The knowledge of this nonlinear elastic parameters paves the way to quantify in vivo the local forces produced by muscle during exercise, contraction or movements.

Comparison of ultrasound elastography, magnetic resonance elastography and finite element model to quantify nonlinear shear modulus.

Objective. The aim of this study is to validate the estimation of the nonlinear shear modulus (A) from the acoustoelasticity theory with two experimental methods, ultrasound (US) elastography and magnetic resonance elastography (MRE), and a finite element method.Approach. Experiments were performed on agar (2%)-gelatin (8%) phantom considered as homogeneous, elastic and isotropic. Two specific setups were built to ensure a uniaxial stress step by step on the phantom, one for US and a nonmagnetic version for MRE. The stress was controlled identically in both imaging techniques, with a water tank placed on the top of the phantom and filled with increasing masses of water during the experiment. In US, the supersonic shear wave elastography was implemented on an ultrafast US device, driving a 6 MHz linear array to measure shear wave speed. In MRE, a gradient-echo sequence was used in which the three spatial directions of a 40 Hz continuous wave displacement generated with an external driver were encoded successively. Numerically, a finite element method was developed to simulate the propagation of the shear wave in a uniaxially stressed soft medium.Main results. Similar shear moduli were estimated at zero stress using experimental methods,µ0US= 12.3 ± 0.3 kPa andµ0MRE= 11.5 ± 0.7 kPa. Numerical simulations were set with a shear modulus of 12 kPa and the resulting nonlinear shear modulus was found to be -58.1 ± 0.7 kPa. A very good agreement between the finite element model and the experimental models (AUS= -58.9 ± 9.9 kPa andAMRE= -52.8 ± 6.5 kPa) was obtained.Significance. These results show the validity of such nonlinear shear modulus measurement quantification in shear wave elastography. This work paves the way to develop nonlinear elastography technique to get a new biomarker for medical diagnosis.

Resistivity index mapping in Kidney based on ultrasensitive Pulsed-Wave Doppler and automatic spectrogram envelope detection.

In recent years, ultrasensitive Pulsed-Wave Doppler (uPWD) ultrasound (US) has emerged as an alternative imaging approach for microcirculation imaging and as a complementary tool to other imaging modalities, such as positron emission tomography (PET). uPWD is based on the acquisition of a large set of highly spatiotemporally coherent frames, which allows high-quality images of a wide field of view to be obtained. In addition, these acquired frames allow calculation of the resistivity index (RI) of the pulsatile flow detected over the entire field of view, which is of great interest to clinicians, for example, in monitoring the transplanted kidney course. This work aims to develop and evaluate a method to automatically obtain an RI map of the kidney based on the uPWD approach. The effect of time gain compensation (TGC) on the visualization of vascularization and aliasing on the blood flow frequency response, was also assessed. A pilot study conducted in patients referred for renal transplant Doppler examination showed that the proposed method provided relative errors of about 15% for RI measurements with respect to conventional pulsed-wave (PW) Doppler.

Multi-layered adaptive neoangiogenesis Intra-Operative quantification (MANIOQ).

Quantification of vascularization volume can provide valuable information for diagnosis and prognosis in vascular pathologies. It can be adapted to inform the surgical management of gliomas, aggressive brain tumors characterized by exuberant sprouting of new blood vessels (neoangiogenesis). Filtered ultrafast Doppler data can provide two main parameters: vascularization index (VI) and fractional moving blood volume (FMBV) that clinically reflect tumor micro vascularization. Current protocols lack robust, automatic, and repeatable filtering methods. We present a filtrating method called Multi-layered Adaptive Neoangiogenesis Intra-Operative Quantification (MANIOQ). First, an adaptive clutter filtering is implemented, based on singular value decomposition (SVD) and hierarchical clustering. Second a method for noise equalization is applied, based on the subtraction of a weighted noise profile. Finally, an in vivo analysis of the periphery of the B-mode hyper signal area allows to measure the vascular infiltration extent of the brain tumors. Ninety ultrasound acquisitions were processed from 23 patients. Compared to reference methods in the literature, MANIOQ provides a more robust tissue filtering, and noise equalization allows for the first time to keep axial and lateral gain compensation (TGC and LGC). MANIOQ opens the way to an intra-operative clinical analysis of gliomas micro vascularization.

Contractile properties are less affected at long than short muscle length after eccentric exercise.

PURPOSE: The aim of the present study was to investigate whether the electrically evoked muscle responses are differently affected over time by the knee joint angle after an exercise-induced muscle damage (EIMD). We hypothesized that low-frequency-evoked responses would be less affected at long than short muscle length, and that mechanisms located within the muscle and tendinous tissues would be involved. METHODS: Fifteen males performed 45 min loaded downhill walking (DW) exercise. Maximal voluntary contraction torque (MVC), optimal angle for torque production, voluntary activation level (VAL), twitch, doublet at 10 and 100 Hz (Db10 and Db100, respectively), rate of torque development (RTD), post-activation potentiation (PAP), muscle shear elastic modulus (µ) and aponeurosis stiffness were assessed before, after, and 4, 24, 48, 72 and 168 h after the exercise at a knee angle of 40°, 90° and 120° (0°: full extension). RESULTS: MVC, VAL and Db100 were similarly decreased across joint angles after the DW and optimal angle was not affected. Twitch, Db10, Db10/Db100, PAP and RTD were less affected and muscle µ more increased at long than short muscle lengths (p < 0.05), especially during the first 24 h after the DW exercise. CONCLUSION: Low-frequency-evoked responses were more preserved at long than short muscle length the first 24 h after the DW exercise, suggesting that joint angle should be taken into account to assess muscular alterations after EIMD. This length-dependence could be associated to the higher sensitivity to Ca2+ and the higher increase in muscle stiffness at long than short muscle length.

Active Heat Acclimation Does Not Alter Muscle-Tendon Unit Properties.

PURPOSE: Heat acclimation (HA) is recommended before competing in hot and humid conditions. HA has also been recently suggested to increase muscle strength, but its effects on human's muscle and tendon mechanical properties are not yet fully understood. This study investigated the effect of active HA on gastrocnemius medialis (GM) muscle-tendon properties. METHODS: Thirty recreationally active participants performed 13 low-intensity cycling sessions, distributed over a 17-d period in hot (HA = ~38°C, ~58% relative humidity; n = 15) or in temperate environment (CON = ~23°C, ~35% relative humidity; n = 15). Mechanical data and high-frame rate ultrasound images were collected during electrically evoked and voluntary contractions pre- and postintervention. Shear modulus was measured at rest in GM, and vertical jump performance was assessed. RESULTS: Core temperature decreased from the first to the last session in HA (-0.4°C ± 0.3°C; P = 0.015), while sweat rate increased (+0.4 ± 0.3 L·h -1 ; P = 0.010), suggesting effective HA, whereas no changes were observed in CON (both P ≥ 0.877). Heart rate was higher in HA versus CON and decreased throughout intervention in groups (both P ≤ 0.008), without an interaction effect ( P = 0.733). Muscle-tendon unit properties (i.e., maximal and explosive isometric torque production, contractile properties, voluntary activation, joint and fascicular force-velocity relationship, passive muscle, and active tendon stiffness) and vertical jump performance did not show training ( P ≥ 0.067) or group-training interaction ( P ≥ 0.232) effects. CONCLUSIONS: Effective active HA does not alter muscle-tendon properties. Preparing hot and humid conditions with active HA can be envisaged in all sporting disciplines without the risk of impairing muscle performance.

Investigation of the relationship between tensile viscoelasticity and unloaded ultrasound shear wave measurements in ex vivo tendon.

Mechanical properties of biological tissues are of key importance for proper function and in situ methods for mechanical characterization are sought after in the context of both medical diagnosis as well as understanding of pathophysiological processes. Shear wave elastography (SWE) and accompanying physical modelling methods provide valid estimates of stiffness in quasi-linear viscoelastic, isotropic tissue but suffer from limitations in assessing non-linear viscoelastic or anisotropic material, such as tendon. Indeed, mathematical modelling predicts the longitudinal shear wave velocity to be unaffected by the tensile but rather the shear viscoelasticity. Here, we employ a heuristic experimental testing approach to the problem to assess the most important potential confounders, namely tendon mass density and diameter, and to investigate associations between tendon tensile viscoelasticity with shear wave descriptors. Small oscillatory testing of animal flexor tendons at two baseline stress levels over a large frequency range comprehensively characterized tensile viscoelastic behavior. A broad set of shear wave descriptors was retrieved on the unloaded tendon based on high frame-rate plane wave ultrasound after applying an acoustic deformation impulse. Tensile modulus and strain energy dissipation increased logarithmically and linearly, respectively, with the frequency of the applied strain. Shear wave descriptors were mostly unaffected by tendon diameter but were highly sensitive to tendon mass density. Shear wave group and phase velocity showed no association with tensile elasticity or strain rate-stiffening but did show an association with tensile strain energy dissipation. The longitudinal shear wave velocity may not characterize tensile elasticity but rather tensile viscous properties of transversely isotropic collagenous tissues.

Transcranial ultrasound simulations: A review.

Transcranial ultrasound is more and more used for therapy and imaging of the brain. However, the skull is a highly attenuating and aberrating medium, with different structures and acoustic properties among samples and even within a sample. Thus, case-specific simulations are needed to perform transcranial focused ultrasound interventions safely. In this article, we provide a review of the different methods used to model the skull and to simulate ultrasound propagation through it.

Mechano-biochemical marine stimulation of inversion, gastrulation, and endomesoderm specification in multicellular Eukaryota.

The evolutionary emergence of the primitive gut in Metazoa is one of the decisive events that conditioned the major evolutionary transition, leading to the origin of animal development. It is thought to have been induced by the specification of the endomesoderm (EM) into the multicellular tissue and its invagination (i.e., gastrulation). However, the biochemical signals underlying the evolutionary emergence of EM specification and gastrulation remain unknown. Herein, we find that hydrodynamic mechanical strains, reminiscent of soft marine flow, trigger active tissue invagination/gastrulation or curvature reversal via a Myo-II-dependent mechanotransductive process in both the metazoan Nematostella vectensis (cnidaria) and the multicellular choanoflagellate Choanoeca flexa. In the latter, our data suggest that the curvature reversal is associated with a sensory-behavioral feeding response. Additionally, like in bilaterian animals, gastrulation in the cnidarian Nematostella vectensis is shown to participate in the biochemical specification of the EM through mechanical activation of the ß-catenin pathway via the phosphorylation of Y654-ßcatenin. Choanoflagellates are considered the closest living relative to metazoans, and the common ancestor of choanoflagellates and metazoans dates back at least 700 million years. Therefore, the present findings using these evolutionarily distant species suggest that the primitive emergence of the gut in Metazoa may have been initiated in response to marine mechanical stress already in multicellular pre-Metazoa. Then, the evolutionary transition may have been achieved by specifying the EM via a mechanosensitive Y654-ßcatenin dependent mechanism, which appeared during early Metazoa evolution and is specifically conserved in all animals.

Anisotropy in ultrasound shear wave elastography: An add-on to muscles characterization.

Ultrasound shear wave elastography was developed the past decade, bringing new stiffness biomarker in clinical practice. This biomarker reveals to be of primarily importance for the diagnosis of breast cancer or liver fibrosis. In muscle this biomarker become much more complex due to the nature of the muscle itself: an anisotropic medium. In this manuscript we depict the underlying theory of propagating waves in such anisotropic medium. Then we present the available methods that can consider and quantify this parameter. Advantages and drawbacks are discussed to open the way to imagine new methods that can free this biomarker in a daily clinical practice.

Poor Correlation between Diaphragm Thickening Fraction and Transdiaphragmatic Pressure in Mechanically Ventilated Patients and Healthy Subjects.

BACKGROUND: The relationship between the diaphragm thickening fraction and the transdiaphragmatic pressure, the reference method to evaluate the diaphragm function, has not been clearly established. This study investigated the global and intraindividual relationship between the thickening fraction of the diaphragm and the transdiaphragmatic pressure. The authors hypothesized that the diaphragm thickening fraction would be positively and significantly correlated to the transdiaphragmatic pressure, in both healthy participants and ventilated patients. METHODS: Fourteen healthy individuals and 25 mechanically ventilated patients (enrolled in two previous physiologic investigations) participated in the current study. The zone of apposition of the right hemidiaphragm was imaged simultaneously to transdiaphragmatic pressure recording within different breathing conditions, i.e., external inspiratory threshold loading in healthy individuals and various pressure support settings in patients. A blinded offline breath-by-breath analysis synchronously computed the changes in transdiaphragmatic pressure, the diaphragm pressure-time product, and diaphragm thickening fraction. Global and intraindividual relationships between variables were assessed. RESULTS: In healthy subjects, both changes in transdiaphragmatic pressure and diaphragm pressure-time product were moderately correlated to diaphragm thickening fraction (repeated measures correlation = 0.40, P < 0.0001; and repeated measures correlation = 0.38, P < 0.0001, respectively). In mechanically ventilated patients, changes in transdiaphragmatic pressure and thickening fraction were weakly correlated (repeated measures correlation = 0.11, P = 0.008), while diaphragm pressure-time product and thickening fraction were not (repeated measures correlation = 0.04, P = 0.396). Individually, changes in transdiaphragmatic pressure and thickening fraction were significantly correlated in 8 of 14 healthy subjects (ρ = 0.30 to 0.85, all P < 0.05) and in 2 of 25 mechanically ventilated patients (ρ = 0.47 to 0.64, all P < 0.05). Diaphragm pressure-time product and thickening fraction correlated in 8 of 14 healthy subjects (ρ = 0.41 to 0.82, all P < 0.02) and in 2 of 25 mechanically ventilated patients (ρ = 0.63 to 0.66, all P < 0.01). CONCLUSIONS: Overall, diaphragm function as assessed with transdiaphragmatic pressure was weakly related to diaphragm thickening fraction. The diaphragm thickening fraction should not be used in healthy subjects or ventilated patients when changes in diaphragm function are evaluated.

Acousto-elasticity of transversely isotropic incompressible soft tissues: characterization of skeletal striated muscle.

Using shear wave elastography, we measure the changes in the wave speed with the stress produced by a striated muscle during isometric voluntary contraction. To isolate the behaviour of an individual muscle from complementary or antagonistic actions of adjacent muscles, we select theflexor digiti minimimuscle, whose sole function is to extend the little finger. To link the wave speed to the stiffness, we develop an acousto-elastic theory for shear waves in homogeneous, transversely isotropic, incompressible solids subject to an uniaxial stress. We then provide measurements of the apparent shear elastic modulus along, and transversely to, the fibre axis for six healthy human volunteers of different age and sex. The results display a great variety across the six subjects. We find that the slope of the apparent shear elastic modulus along the fibre direction changes inversely to the maximum voluntary contraction (MVC) produced by the volunteer. We propose an interpretation of our results by introducing the S (slow) or F (fast) nature of the fibres, which harden the muscle differently and accordingly, produce different MVCs. A natural follow-up on this study is to apply the method to patients with musculoskeletal disorders or neurodegenerative diseases.

Tumor Solid Stress: Assessment with MR Elastography under Compression of Patient-Derived Hepatocellular Carcinomas and Cholangiocarcinomas Xenografted in Mice.

Malignant tumors have abnormal biomechanical characteristics, including high viscoelasticity, solid stress, and interstitial fluid pressure. Magnetic resonance (MR) elastography is increasingly used to non-invasively assess tissue viscoelasticity. However, solid stress and interstitial fluid pressure measurements are performed with invasive methods. We studied the feasibility and potential role of MR elastography at basal state and under controlled compression in assessing altered biomechanical features of malignant liver tumors. MR elastography was performed in mice with patient-derived, subcutaneously xenografted hepatocellular carcinomas or cholangiocarcinomas to measure the basal viscoelasticity and the compression stiffening rate, which corresponds to the slope of elasticity versus applied compression. MR elastography measurements were correlated with invasive pressure measurements and digital histological readings. Significant differences in MR elastography parameters, pressure, and histological measurements were observed between tumor models. In multivariate analysis, collagen content and interstitial fluid pressure were determinants of basal viscoelasticity, whereas solid stress, in addition to collagen content, cellularity, and tumor type, was an independent determinant of compression stiffening rate. Compression stiffening rate had high AUC (0.87 ± 0.08) for determining elevated solid stress, whereas basal elasticity had high AUC for tumor collagen content (AUC: 0.86 ± 0.08). Our results suggest that MR elastography compression stiffening rate, in contrast to basal viscoelasticity, is a potential marker of solid stress in malignant liver tumors.

Diagnostic Accuracy of Four Levels of Manual Compression Applied in Supersonic Shear Wave Elastography of the Breast.

PURPOSE: To investigate the diagnostic accuracy of applying four levels of manual pressure in Shear Wave Elastography (SWE) of the breast and to assess inter-rater reliability. MATERIALS AND METHODS: Single-center prospective preliminary study including patients receiving ultrasound examination of breast lesions as part of routine clinical practice. SWE was performed on 60 breast masses (26 benign and 34 malignant) in 54 patients by a breast fellowship trained radiologist. Stiffness values were compared between benign and malignant masses at four levels of manual compression: none, mild, moderate, and marked. Accuracy of SWE was assessed using receiving operating characteristics analysis at each level. In 18 patients, a second radiologist repeated the SWE acquisitions to evaluate reproducibility. Reproducibility was assessed using intraclass correlation coefficient. RESULTS: Without compression, we observed no significant difference in stiffness (p > 0.99) between benign and malignant lesions, and SWE demonstrated low accuracy (area under the curve = 0.64). Stiffness was higher in malignant lesions at all levels of compression (p < 0.001). SWE demonstrated good accuracy at all three levels of compression (from area under the curve = 0.71 to 0.84 across Emax and Emean), with high interobserver agreement. CONCLUSION: This preliminary study suggests that not using compression during SWE for breast lesion characterization offers suboptimal results. On the contrary, application of compression yields high diagnostic performance with good interobserver agreement and, as such, should be included in routine clinical practice.

Acoustoelasticity in transversely isotropic soft tissues: Quantification of muscle nonlinear elasticity.

Recent developments in the field of elastography aim at developing the quantification of new mechanical properties of tissues, that are complementary to the shear modulus, which is characteristic of the linear elastic properties of a quasi-incompressible medium. In this context, measurement of the elastic nonlinearity of tissues was recently proposed based on acoustoelasticity. Up to now, most of the experimental applications of acoustoelasticity theory using Landau formalism in human tissues have assumed isotropy. However, this strong hypothesis does not hold in all human tissues, such as muscles that are generally considered as transversely isotropic (TI). In this work, after reviewing the constraints imposed by TI symmetry on the linear and nonlinear elastic properties of TI media, the acoustoelasticity theory in TI incompressible media is developed and implemented experimentally on a TI polyvinyl alcohol phantom and on ex vivo muscular tissues. Based on this theory and on the evolutions of the shear wave speed, with respect to uniaxial static stress, the nonlinear elastic parameter A is experimentally quantified. The estimations of A in ex vivo bovine and porcine muscles are on the order of hundreds of kPa. This work paves the way for more thorough muscle mechanical properties characterization as well as for the development of a potential new biomarker.

Challenges and Perspectives of the Hybridization of PET with Functional MRI or Ultrasound for Neuroimaging.

Hybridization of positron emission tomography (PET) with other functional neuroimaging techniques such as functional magnetic resonance imaging (fMRI) or functional ultrasound (fUS) still raises technical and methodological challenges. Beyond the co-registration of anatomical images with functional data, development of hybrid imaging systems has paved the way for a large field of research based on the concept of bimodal functional neuroimaging such as PET/fMRI. In this framework, comparison of respective performances of brain PET and fUS suggests complementarity and great potential of hybrid PET/fUS for preclinical neuroimaging. Hybridization of functional neuroimaging techniques first offers opportunities to validate or improve measurement made by each modality. Future research may propose and validate hybrid parameters that quantitatively connect the brain molecular environment and the neuro-vascular coupling, which may improve our understanding of brain function in health and disease, with perspectives in neuroscience and neuropharmacology. In the coming years, cross-fertilization of neuroimaging communities and training of young researchers on multiple imaging modalities may foster the development of hybrid neuroimaging protocols that will take the full potential and the limitations of each modality into account.

Ultrasound shear wave elastography for assessing diaphragm function in mechanically ventilated patients: a breath-by-breath analysis.

BACKGROUND: Diaphragm dysfunction is highly prevalent in mechanically ventilated patients. Recent work showed that changes in diaphragm shear modulus (ΔSMdi) assessed using ultrasound shear wave elastography (SWE) are strongly related to changes in Pdi (ΔPdi) in healthy subjects. The aims of this study were to investigate the relationship between ΔSMdi and ΔPdi in mechanically ventilated patients, and whether ΔSMdi is responsive to change in respiratory load when varying the ventilator settings. METHODS: A prospective, monocentric study was conducted in a 15-bed ICU. Patients were included if they met the readiness-to-wean criteria. Pdi was continuously monitored using a double-balloon feeding catheter orally introduced. The zone of apposition of the right hemidiaphragm was imaged using a linear transducer (SL10-2, Aixplorer, Supersonic Imagine, France). Ultrasound recordings were performed under various pressure support settings and during a spontaneous breathing trial (SBT). A breath-by-breath analysis was performed, allowing the direct comparison between ΔPdi and ΔSMdi. Pearson's correlation coefficients (r) were used to investigate within-individual relationships between variables, and repeated measure correlations (R) were used for determining overall relationships between variables. Linear mixed models were used to compare breathing indices across the conditions of ventilation. RESULTS: Thirty patients were included and 930 respiratory cycles were analyzed. Twenty-five were considered for the analysis. A significant correlation was found between ΔPdi and ΔSMdi (R = 0.45, 95% CIs [0.35 0.54], p < 0.001). Individual correlation displays a significant correlation in 8 patients out of 25 (r = 0.55-0.86, all p < 0.05, versus r = - 0.43-0.52, all p > 0.06). Changing the condition of ventilation similarly affected ΔPdi and ΔSMdi. Patients in which ΔPdi-ΔSMdi correlation was non-significant had a faster respiratory rate as compared to that of patient with a significant ΔPdi-ΔSMdi relationship (median (Q1-Q3), 25 (18-33) vs. 21 (15-26) breaths.min-1, respectively). CONCLUSIONS: We demonstrate that ultrasound SWE may be a promising surrogate to Pdi in mechanically ventilated patients. Respiratory rate appears to negatively impact SMdi measurement. Technological developments are needed to generalize this method in tachypneic patients. TRIAL REGISTRATION: NCT03832231 .

Evaluation of capacitive micromachined ultrasonic transducers for passive monitoring of microbubble-assisted ultrasound therapies.

Passive cavitation detection can be performed to monitor microbubble activity during brain therapy. Microbubbles under ultrasound exposure generate a response characterized by multiple nonlinear emissions. Here, the wide bandwidth of capacitive micromachined ultrasonic transducers (CMUTs) was exploited to monitor the microbubble signature through a rat skull and a macaque skull. The intrinsic nonlinearity of the CMUTs was characterized in receive mode. Indeed, undesirable nonlinear components generated by the CMUTs must be minimized as they can mask the microbubble harmonic response. The microbubble signature at harmonic and ultra-harmonic components (0.5-6 MHz) was successfully extracted through a rat skull using moderate bias voltage.