Detection of Individual Microbubbles by Burst-Wave-Aided Contrast-Enhanced Active Doppler Ultrasonography.

We propose burst-wave-aided, contrast-enhanced, active Doppler ultrasonography for visualizing lymph vessels. This technique forces ultrasound contrast agents (UCAs) to move using the acoustic radiation force induced by burst waves with low amplitude while suppressing their destruction. Using a flow phantom, we measured the average, decrease rate of echo intensity [i.e., pulse intensity integral (PII)], and the velocity of individual contrast agents, which directly affects the performance of imaging and tracking contrast agents under stationary flow conditions. Comparison with pulse-inversion Doppler without exposure to the burst wave demonstrated that the velocity of the contrast agents could be enhanced up to several tens of millimeters per second by the effect of the burst wave, maximizing the echo intensity extracted by a clutter filter. The contrast ratio (CR), defined as the ratio of the contrast echo to the phantom echo outside the channel, did not change appreciably, even when the lower cut-off velocity of the clutter filter was increased up to 10 mm/s. This implies a better robustness against the motion of the tissue. In addition, the performance for detecting contrast agents (i.e., echo intensity) was superior or similar to that of pulse-inversion Doppler, even in undesirable conditions where the flow had a velocity component in the opposite direction to that of the acoustic radiation force. The echo intensity was lower or the same as that in pulse-inversion Doppler, demonstrating the potential for suppressing the destruction of contrast agents and enabling long-term observations. From these results, we expect that the proposed method will be beneficial for visualizing lymph vessels.

Investigation of a method to estimate the average speed of sound using phase variances of element signals for ultrasound compound imaging.

PURPOSE: In the receive beamforming of an ultrasonography system, a B-mode image is reconstructed by assuming an average speed of sound (SoS) as a constant value. In our previous studies, we proposed a method for estimating the average SoS based on the coherence factor (CF) and the reciprocal of phase variances of element signals in delay-and-sum (DAS) beamforming. In this paper, we investigate the accuracy of estimation of the average SoS for compound imaging. METHODS: For this purpose, two numerical simulations were performed with k-Wave software. Also, the estimation methods based on the CF and the reciprocal were applied to in vivo data from the common carotid artery, and B-mode images were reconstructed using the estimated average SoS. RESULTS: In the first numerical simulation using an inhomogeneous phantom, the relationship between the accuracy and the transmission angles for the estimation was investigated, and the root mean squared errors (RMSEs) of estimates obtained based on the CF and the reciprocal of the phase variance were 1.25 ± 0.09, and 0.765 ± 0.17% at the transmission sequence of steering angles of (- 10°, - 5°, 0°, 5°, 10°), respectively. In the second numerical simulation using a cyst phantom, lateral resolutions were improved by reconstructing the image using the estimates obtained using the proposed strategy (reciprocal). By the proposed strategy, improvement of the continuity of the lumen-intima interface in the lateral direction was observed in the in vivo experiment. CONCLUSION: Consequently, the results indicated that the proposed strategy was beneficial for estimation of the average SoS and image reconstruction.

Effect of Clutter Filter in High-Frame-Rate Ultrasonic Backscatter Coefficient Analysis.

High-frame-rate imaging with a clutter filter can clearly visualize blood flow signals and provide more efficient discrimination with tissue signals. In vitro studies using clutter-less phantom and high-frequency ultrasound suggested a possibility of evaluating the red blood cell (RBC) aggregation by analyzing the frequency dependence of the backscatter coefficient (BSC). However, in in vivo applications, clutter filtering is required to visualize echoes from the RBC. This study initially evaluated the effect of the clutter filter for ultrasonic BSC analysis for in vitro and preliminary in vivo data to characterize hemorheology. Coherently compounded plane wave imaging at a frame rate of 2 kHz was carried out in high-frame-rate imaging. Two samples of RBCs suspended by saline and autologous plasma for in vitro data were circulated in two types of flow phantoms without or with clutter signals. The singular value decomposition was applied to suppress the clutter signal in the flow phantom. The BSC was calculated using the reference phantom method, and it was parametrized by spectral slope and mid-band fit (MBF) between 4-12 MHz. The velocity distribution was estimated by the block matching method, and the shear rate was estimated by the least squares approximation of the slope near the wall. Consequently, the spectral slope of the saline sample was always around four (Rayleigh scattering), independently of the shear rate, because the RBCs did not aggregate in the solution. Conversely, the spectral slope of the plasma sample was lower than four at low shear rates but approached four by increasing the shear rate, because the aggregations were presumably dissolved by the high shear rate. Moreover, the MBF of the plasma sample decreased from -36 to -49 dB in both flow phantoms with increasing shear rates, from approximately 10 to 100 s-1. The variation in the spectral slope and MBF in the saline sample was comparable to the results of in vivo cases in healthy human jugular veins when the tissue and blood flow signals could be separated.

Contrast analysis in ultrafast ultrasound blood flow imaging of jugular vein.

PURPOSE: The contrasts of flowing blood in in vitro experiments using porcine blood and in vivo measurements of human jugular veins were analyzed to demonstrate that the hemorheological property was dependent on the shear rate. METHODS: Blood samples (45% hematocrit) suspended in saline or plasma were compared with examine the difference in viscoelasticity. Ultrafast plane-wave imaging at an ultrasonic center frequency of 7.5 MHz was performed on different steady flows in a graphite-agar phantom. Also, in vivo measurement was performed in young, healthy subjects and patients with diabetes. A spatiotemporal matrix of beamformed radio-frequency data was used for the singular value decomposition (SVD) clutter filter. The clutter-filtered B-mode image was calculated as the amplitude envelope normalized at the first frame in the diastolic phase to evaluate contrast. The shear rate was estimated as the velocity gradient perpendicular to the lateral axis. RESULTS: Although nonaggregated erythrocytes at a high shear rate exhibited a low echogenicity, the echogenicity in the plasma sample overall increased due to erythrocyte aggregation at a low shear rate. In addition, the frequency of detection of specular components, defined as components beyond twice the standard deviation of a contrast map obtained from a clutter-filtered B-mode image, increased in the porcine blood at a high shear rate and the venous blood in healthy subjects versus patients with diabetes. CONCLUSION: The possibility of characterizing hemorheological properties dependent on the shear rate and diabetes condition was indicated using ultrafast plane-wave imaging with an SVD-based clutter filter.

Two-Dimensional Wavenumber Analysis Implemented in Ultrasonic Vector Doppler Method with Focused Transmit Beams.

The multi-angle Doppler method was introduced for the estimation of velocity vectors by measuring axial velocities from multiple directions. We have recently reported that the autocorrelation-based velocity vector estimation could be ameliorated significantly by estimating the wavenumbers in two dimensions. Since two-dimensional wavenumber estimation requires a snapshot of an ultrasonic field, the method was first implemented in plane wave imaging. Although plane wave imaging is predominantly useful for examining blood flows at an extremely high temporal resolution, it was reported that the contrast in a B-mode image obtained with a few plane wave emissions was lower than that obtained with focused beams. In this study, the two-dimensional wavenumber analysis was first implemented in a framework with focused transmit beams. The simulations showed that the proposed method achieved an accuracy in velocity estimation comparable to that of the method with plane wave imaging. Furthermore, the performances of the methods implemented in focused beam and plane wave imaging were compared by measuring human common carotid arteries in vivo. Image contrasts were analyzed in normal and clutter-filtered B-mode images. The method with focused beam imaging achieved a better contrast in normal B-mode imaging, and similar velocity magnitudes and angles were obtained by both the methods with focused beam and plane wave imaging. In contrast, the method with plane wave imaging gave a better contrast in a clutter-filtered B-mode image and smaller variances in velocity magnitudes than those with focused beams.

On the Investigation of Autocorrelation-Based Vector Doppler Method With Plane Wave Imaging.

Although color flow imaging is one of the representative applications of the Doppler method, it can estimate only the velocity component in the direction of ultrasonic propagation, that is, the axial velocity component. The vector Doppler method with high-frame-rate plane wave imaging overcomes such a limitation by estimating the blood flow velocity vectors using the axial velocities obtained by emitting plane waves in multiple directions. The autocorrelation technique can be used for the estimation of the axial velocity using the phase shift of an ultrasonic echo signal between two transmit-receive events. The technique also requires the frequency of the received echo signal. Although the center frequency of the emitted ultrasonic signal is commonly used in the estimation of axial velocities, the center frequency should be estimated from the received signals. In this study, a method for the estimation of the center frequency designed particularly for the high-frame-rate plane wave imaging was developed. The proposed method estimates the wavenumbers of the received signal in lateral and vertical directions to estimate the wavenumber in the axial direction, from which the center frequency was estimated. The beam steering angle was also estimated from the wavenumbers in the two directions. The effect of the proposed method was validated in simulations. The absolute bias error (ABE) and root-mean squared error in estimated velocity vectors obtained by plane wave imaging with three beam steering angles (-15°, 0°, and 15°) were reduced from 9.27% and 14.80% to 1.15% and 8.75%, respectively, by the proposed method. The applicability of the proposed method to in vivo measurements was also demonstrated using the in vivo recordings of human common carotid arteries. Physiologically consistent blood flow velocity distributions were obtained with respect to three subjects using the proposed method.

In Vivo Quantitative Ultrasound on Dermis and Hypodermis for Classifying Lymphedema Severity in Humans.

This study investigated the ability of in vivo quantitative ultrasound (QUS) assessment to evaluate lymphedema severity compared with the gold standard method, the International Society of Lymphology (ISL) stage. Ultrasonic measurements were made around the middle thigh (n = 150). Radiofrequency data were acquired using a clinical scanner and 8-MHz linear probe. Envelope statistical analysis was performed using constant false alarm rate processing and homodyned K (HK) distribution. The attenuation coefficient was calculated using the spectral log-difference technique. The backscatter coefficient (BSC) was obtained by the reference phantom method with attenuation compensation according to the attenuation coefficients in the dermis and hypodermis, and then effective scatterer diameter (ESD) and effective acoustic concentration (EAC) were estimated with a Gaussian model. Receiver operating characteristic curves of QUS parameters were obtained using a linear regression model. A single QUS parameter with high area under the curve (AUC) differed between the dermis (ESD and EAC) and hypodermis (HK) parameters. The combinations with ESD and EAC in the dermis, HK parameters in the hypodermis and typical features (dermal thickness and echogenic regions in the hypodermis) improved classification performance between ISL stages 0 and ≥I (AUC = 0.90 with sensitivity of 75% and specificity of 91%) in comparison with ESD and EAC in the dermis (AUC = 0.82) and HK parameters in the hypodermis (AUC = 0.82). In vivo QUS assessment by BSC and envelope statistical analyses can be valuable for non-invasively classifying an extremely early stage of lymphedema, such as ISL stage I, and following its progression.

Statistical Analysis of Ultrasonic Scattered Echoes Enables the Non-invasive Measurement of Temperature Elevations inside Tumor Tissue during Oncological Hyperthermia.

Non-invasive monitoring of temperature elevations inside tumor tissue is imperative for the oncological thermotherapy known as hyperthermia. In the present study, two cancer patients, one with a developing right renal cell carcinoma and the other with pseudomyxoma peritonei, underwent hyperthermia. The two patients were irradiated with radiofrequency current for 40 min during hyperthermia. We report the results of our clinical trial study in which the temperature increases inside the tumor tissues of patients with right renal cell carcinoma and pseudomyxoma peritonei induced by radiofrequency current irradiation for 40 min could be detected by statistical analysis of ultrasonic scattered echoes. The Nakagami shape parameter m varies depending on the temperature of the medium. We calculated the Nakagami shape parameter m by statistical analysis of the ultrasonic echoes scattered from the tumor tissues. The temperature elevations inside the tumor tissues were expressed as increases in brightness on 2-D hot-scale maps of the specific parameter αmod, indicating the absolute values of the percentage changes in m values. In the αmod map for each tumor tissue, the brightness clearly increased with treatment time. In quantitative analysis, the mean values of αmod were calculated. The mean value of αmod for the right renal cell carcinoma increased to 1.35 dB with increasing treatment time, and the mean value of αmod for pseudomyxoma peritonei increased to 1.74 with treatment time. The increase in both αmod brightness and the mean value of αmod implied temperature elevations inside the tumor tissues induced by the radiofrequency current; thus, the acoustic method is promising for monitoring temperature elevations inside tumor tissues during hyperthermia.

Measurement of flow velocity vectors in carotid artery using plane wave imaging with repeated transmit sequence.

PURPOSE: Doppler-based methods are widely used for blood flow imaging in clinical settings. However, they inherently estimate the velocity component only in the axial direction. Therefore, various studies of angle-independent methods have been conducted. The multi-angle Doppler method is one such angle-independent method, in which the velocity vector is estimated using axial velocities obtained from multiple directions by steering an ultrasonic beam. Recently, plane wave imaging, which realizes a very high frame rate of several thousand frames per second, was applied to the multi-angle Doppler method. However, the maximum detectable velocity, i.e., the aliasing limit, was reduced depending on the number of steering angles. In the present study, the feasibility of a specific transmit sequence, namely, the repeated transmit sequence, was examined using the plane-wave multi-angle Doppler method. METHOD: In the repeated transmit sequence, plane waves were emitted to the same direction twice, after which the steering angle was changed. By repeating the same procedure, a pair of beamformed radio-frequency (RF) signals could be obtained under each beam steering angle. By applying the autocorrelation method to each pair of RF signals, the time interval between the RF signals could be kept as the pulse repetition interval (PRI). The feasibility of such a transmit sequence was examined by numerical simulation and in vivo measurement of a human carotid artery. RESULTS: The simulation results showed that the maximum steering angles of over 10 degrees were not feasible with the linear array used in the present study. The feasible maximum steering angle would depend on the element pitch of the probe relative to the ultrasonic wavelength. By limiting the maximum steering angles to 5 and 10 degrees, bias errors were 9.2% and 11.3%, respectively, and root mean squared errors were 21.5% and 16.9%, respectively. Also, flow velocity vectors in a human carotid artery could be visualized with the proposed method. CONCLUSION: The multi-angle Doppler method was implemented in plane wave imaging with the repeated transmit sequence, and the proposed method was shown to be feasible through numerical simulation and in vivo measurement of a carotid artery.

Noninvasive, objective evaluation of lower extremity lymphedema severity using shear wave elastography: A preliminary study.

BACKGROUND: Increased skin and subcutaneous tissue stiffness in patients with early-stage lymphedema has been reported. The purpose of this study was to examine the use of shear wave elastography (SWE) for evaluating lower extremity lymphedema (LEL). METHODS: For 10 lower extremities of normal controls and 72 limbs of patients with gynecological cancer whose lymphatic function was categorized into six stages based on the range of dermal backflow (DBF) observed in indocyanine green (ICG) lymphography, SWE was performed and shear wave velocity (SWV) of the dermis and three layers of subcutaneous tissue at the thigh and calf were recorded. Twenty-five patients underwent thigh tissue histological and dermal thickness examinations. RESULTS: The strongest correlation between the ICG DBF stage and SWV during SWE was observed on the dermal layer of the thigh (p < 0.01, R = 0.67). There was a significant correlation between the dermal thickness of the thigh and the ICG DBF stage (p < 0.01, R = 0.87) and also between the dermal thickness of the thigh and SWV (p < 0.01, R = 0.73). CONCLUSION: Noninvasive, objective evaluation of LEL severity using SWE was well correlated with lymphatic function as determined by ICG lymphography. The DBF changes in the dermis of the thigh best reflected the changes in lymphatic function. Dermal thickness variations may partially account for differences in SWV.

A study on understanding the physical mechanism of change in ultrasonic envelope statistical property during temperature elevation.

PURPOSE: Our previous studies demonstrate that the variation in ultrasonic envelope statistics is correlated with the temperature change inside scattering media. This variation is identified as the change in the scatterer structure during thermal expansion or contraction. However, no specific evidence has been verified to date. This study numerically reproduces the change in the scatterer distribution during thermal expansion or contraction using finite element simulations and also investigates how the situation is altered by different material properties. METHODS: The material properties of a linear elastic solid depend on the thermal expansion coefficient, thermal conductivity, specific heat, and initial scatterer number density. Three-dimensional displacements, calculated in the simulation, were sequentially used to update the positions of the randomly distributed scatterers. Ultrasound signals from the scatterer distribution were generated by simulating a 7.5-MHz linear array transducer whose specifications were the same as those in the experimental measurements of several phantoms and excised porcine livers. To represent the change in the envelope statistical feature, the absolute value of the ratio change in the logarithmic Nakagami (NA) parameter, Δ m , at each time was calculated as a value normalized with the initial NA parameter. RESULTS: The change in the scatterer number density relates to the volume change during temperature elevation. The magnitude of the Δ m shift against the temperature change increases depending on the higher thermal expansion coefficient. In contrast, the relationship between Δ m and the scatterer number density is similar with any material property. Additionally, the changes in Δ m obtained by several experimental phantoms with low to high scatterer number densities are comparable with the numerical simulation results. CONCLUSIONS: The change in Δ m is indirectly related to the change in the scatterer number density owing to the volume change during thermal expansion or contraction.

Investigation of feasibility of singular value decomposition clutter filter in plane wave imaging with packet transmission sequence.

PURPOSE: Assessment of blood flow is an important function in diagnostic ultrasound imaging. Color flow imaging is one such method widely used in the clinical setting. Since autocorrelation suffers from aliasing, the time interval between successive transmissions of ultrasonic pulses should be as short as possible. For this purpose, a specific transmit-receive sequence, namely, packet transmission, is widely used in color flow imaging. Also, plane wave imaging recently introduced to ultrasound imaging significantly contributes to improvement of the temporal resolution. Furthermore, a singular value decomposition (SVD) clutter filter reportedly outperforms a conventional clutter filter. In the present study, the feasibility of the SVD clutter filter in plane wave imaging with the packet transmission sequence was investigated. METHOD: In the present study, the packet transmission sequence was implemented in plane wave imaging by sending plane waves multiple times in the same direction before changing the steering angle. In the first strategy, like conventional color flow imaging with line-by-line acquisition using a focused transmit beam, a clutter filter was applied to ultrasonic radio-frequency (RF) signals in each packet. In the second strategy, the number of transmissions per packet was set at two, and a clutter filter was applied to RF signals obtained from the first or second transmission in different packets. RESULTS: The in vivo experimental results on a human carotid artery showed that the second strategy with an SVD filter realized significantly better performance than the first strategy with a polynomial regression filter used as a conventional filter. CONCLUSION: An SVD clutter filter was feasible in plane wave imaging with the packet transmission sequence, and the performance was improved by limiting the number of transmissions per packet to two.

Frequency dependence of attenuation and backscatter coefficient of ex vivo human lymphedema dermis.

PURPOSE: Radio-frequency (RF) signals from the most dominant scatterer in a dermis, i.e., collagen fibers, are collected as backscattered signals. We aim to confirm the frequency dependence of the spatial distribution of features in ultrasound images, as well as the attenuation coefficient (AC) and backscatter coefficient (BSC) of skin tissue without [LE (-)] and with lymphedema [LE (+)]. METHODS: Measurement samples (n = 13) were excised from human skin tissue with LE (-) and middle severity LE (+). A laboratory-made scanner and single-element concave transducers (range 9-47 MHz) were used to measure RF data. A localized AC was computed from the normalized power spectrum using the linear least squares technique. The reflector method and compensation technique of the attenuation of tissue were applied to calculate the BSC. In addition, effective scatterer diameter (ESD), effective acoustic concentration (EAC), and integrated BSC (IBS) were calculated from the BSC as the benchmark to differentiate LE (-) and LE (+) tissues. RESULTS: High-frequency ultrasound displayed different echogenicity and texture compared between LE (-) and LE (+) in all transducers. The AC for LE (-) (0.22 dB/mm/MHz) and LE (+) (0.29 dB/mm/MHz) was comparable. BSC in LE (-) and LE (+) increased linearly with each transducer. The difference of intercept of the BSC between LE (-) and LE (+) indicated that both EAC and IBS of LE (+) were higher than that of LE (-). In contrast, ESD correlated with the slope of the BSC demonstrated the same tendency for both LE (-) and LE (+). These tendencies appeared for each transducer independent of the frequency bandwidth. CONCLUSION: Frequency independence of AC and BSC in LE (-) and LE (+) was confirmed. Several 9- to 19-MHz ultrasound beams are sufficient for BSC analysis to discriminate LE (-) and LE (+) in terms of the penetration depth of the ultrasound.

Validation of differences in backscatter coefficients among four ultrasound scanners with different beamforming methods.

PURPOSE: The backscatter coefficient (BSC) indicates the absolute scatterer property of a material, independently of clinicians and system settings. Our study verified that the BSC differed among the scanners, transducers, and beamforming methods used for quantitative ultrasound analyses of biological tissues. METHODS: Measurements were performed on four tissue-mimicking homogeneous phantoms containing spherical scatterers with mean diameters of 20 and 30 µm prepared at concentrations of 0.5 and 2.0 wt%, respectively. The BSCs in the different systems were compared using ultrasound scanners with two single-element transducers and five linear high- or low-frequency probes. The beamforming methods were line-by-line formation using focused imaging (FI) and parallel beam formation using plane wave imaging (PWI). The BSC of each system was calculated by the reference phantom method. The mean deviation from the theoretical BSC computed by the Faran model was analyzed as the benchmark validation of the calculated BSC. RESULTS: The BSCs calculated in systems with different properties and beamforming methods well concurred with the theoretical BSC. The mean deviation was below ± 2.8 dB on average, and within the approximate standard deviation (± 2.2 dB at most) in all cases. These variations agreed with a previous study in which the largest error among four different scanners with FI beamforming was 3.5 dB. CONCLUSION: The BSC in PWI was equivalent to those in the other systems and to those of FI beamforming. This result indicates the possibility of ultra-high frame-rate BSC analysis using PWI.

Ultrasound assessment of translation of microbubbles driven by acoustic radiation force in a channel filled with stationary fluid.

In this report, a method is proposed to quantify the translation of ultrasound contrast agent (UCA) microbubbles driven by acoustic radiation for the detection of channels filled with stationary fluid. The authors subjected UCA microbubbles in a channel with diameters of 0.1 and 0.5 mm to ultrasound pulses with a center frequency of 14.4 MHz. The translational velocity of the UCA microbubbles increased with the sound pressure and pulse repetition frequency (PRF) of the transmitted ultrasound. The mean translational velocity reached 0.75 mm/s at a negative peak sound pressure of 2.76 MPa and a PRF of 2 kHz. This trend agreed with the theoretical prediction, which indicated that the translational velocity was proportional to the square of the sound pressure and the PRF. Furthermore, an experiment was carried out with a phantom that mimics tissue and found that the proposed method aided in detection of the channel, even in the case of a low contrast-echo to tissue-echo ratio. The authors expect to develop the proposed method into a technique for detecting lymph vessels.

Cerebellar folding is initiated by mechanical constraints on a fluid-like layer without a cellular pre-pattern.

Models based in differential expansion of elastic material, axonal constraints, directed growth, or multi-phasic combinations have been proposed to explain brain folding. However, the cellular and physical processes present during folding have not been defined. We used the murine cerebellum to challenge folding models with in vivo data. We show that at folding initiation differential expansion is created by the outer layer of proliferating progenitors expanding faster than the core. However, the stiffness differential, compressive forces, and emergent thickness variations required by elastic material models are not present. We find that folding occurs without an obvious cellular pre-pattern, that the outer layer expansion is uniform and fluid-like, and that the cerebellum is under radial and circumferential constraints. Lastly, we find that a multi-phase model incorporating differential expansion of a fluid outer layer and radial and circumferential constraints approximates the in vivo shape evolution observed during initiation of cerebellar folding.