Vessel Rupture Thresholds for Vessel-Bubble Interactions Using an Earthworm Vasculature Model.

OBJECTIVE: Intravenous microbubble oscillation in the presence of ultrasound has the potential to yield a wide range of therapeutic benefits. However, the likelihood of vessel damage caused by mechanical effects has not been quantified as a function of the numerous important parameters in therapeutic ultrasound procedures. In this study, we examined the effects of microbubbles injected into the vasculature of the earthworm. It was found that the elastic properties of earthworm blood vessels are similar to those of arteries in older humans, and that earthworms are well suited to the large number of experiments necessary to investigate safety of procedures involving microbubble oscillation in sonicated vessels. METHODS: Microbubbles were infused into earthworm vessels, and the rupture time during sonication was recorded as a function of ultrasound frequency, pulse repetition frequency and acoustic pressure. DISCUSSION: A modified mechanical index (MMI) was defined that successfully captured the trends in rupture probability and rupture time for the different parameter values, creating a database of vessel rupture thresholds. In the absence of bubbles, the product of MMI squared and rupture time was approximately constant, indicating a possible radiation-force effect. CONCLUSION: The MMI was an effective correlating parameter in the presence of bubbles, though the mathematical dependence is not yet apparent. The results of the study are expected to be valuable in designing more refined studies in vertebrate models, as well as informing computational models.

Earthworm, Lumbricus Terrestris: A Novel Microinjection Vasculature In vivo Invertebrate Model.

Although vertebrates are indispensable to biomedical research, studies are often limited by factors such as cost, lengthy internal review, and ethical considerations. We present the earthworm as an alternative, low-cost, invertebrate applicable to certain preliminary vasculature studies. Due to the surgical availability of the earthworm's dorsal vessels, ventral vessels, and five pairs of pseudo hearts, earthworms are readily accessible, offer low-cost maintenance, and require administration of only small doses of a given compound. The earthworm model provides a simple closed vascular circulatory system with a hemoglobin structure similar to human blood. A protocol is provided for anaesthetizing the earthworms and performing surgical incisions to expose relevant blood vessels. Micropipettes for compound administration are formed by heating and pulling glass with a pipette puller and using a beveling system to create a micron-scale fine needle tip. The tips are then used with a micropositioner and microinjector to inject arbitrary compounds into the vascular system of an earthworm, repeatably, with the availability of large sample sizes and small compound volumes. Details on the intricacies of injection procedure are provided. The small vessel size of the earthworm is challenging, particularly in the case of the ventral vessel; however, mastery of the techniques presented offers high repeatability as a low-cost solution, making studies of very large sample size practical.

Passive Acoustic Mapping with the Angular Spectrum Method.

In the present proof of principle study, we evaluated the homogenous angular spectrum method for passive acoustic mapping (AS-PAM) of microbubble oscillations using simulated and experimental data. In the simulated data we assessed the ability of AS-PAM to form 3D maps of a single and multiple point sources. Then, in the two dimensional limit, we compared the 2D maps from AS-PAM with alternative frequency and time domain passive acoustic mapping (FD- and TD-PAM) approaches. Finally, we assessed the ability of AS-PAM to visualize microbubble activity in vivo with data obtained during 8 different experiments of FUS-induced blood-brain barrier disruption in 3 nonhuman primates, using a clinical MR-guided FUS system. Our in silico results demonstrate AS-PAM can be used to perform 3D passive acoustic mapping. 2D AS-PAM as compared to FD- PAM and TD-PAM is 10 and 200 times faster respectively and has similar sensitivity, resolution, and localization accuracy, even when the noise was 10-fold higher than the signal. In-vivo, the AS-PAM reconstructions of emissions at frequency bands pertinent to the different types of microbubble oscillations were also found to be more sensitive than TD-PAM. AS-PAM of harmonic-only components predicted safe blood-brain barrier disruption, whereas AS-PAM of broadband emissions correctly identified MR-evident tissue damage. The disparity (3.2 mm) in the location of the cavitation activity between the three methods was within their resolution limits. These data clearly demonstrate that AS-PAM is a sensitive and fast approach for PAM, thus providing a clinically relevant method to guide therapeutic ultrasound procedures.

Investigation of the correlation between diffuse infrared and ultrasound for transcranial ultrasound.

Over the past two decades the feasibility for using transcranial ultrasound as both a therapeutic and diagnostic tool has been established. Various aberration-correction techniques have been proposed to achieve transcranial focusing, including CT-derived model based corrections, ultrasound-derived model based corrections, magnetic resonance acoustic radiation force (MR-ARFI) techniques, and techniques involving the invasive introduction of an acoustic source or receiver into the brain. Here, we investigate the correlation between transcranial infrared light (IR) and transcranial ultrasound, where we examine whether IR could be an indicator of any of the key acoustic properties that affect transcranial transmission (signal attenuation, speed of sound, and bone density). Nine human skull samples were utilized in the study. The interior of each sample was illuminated over its inner surface using a diffuse light source. Light transmitted to the outer surface was detected by a 3-mm diameter 940-nm infrared sensor. Acoustic measurements were likewise obtained in a water tank using a 12.7-mm diameter 1-MHz source and a needle hydrophone receiver. Results reveal a positive correlation between the acoustic time-of-flight and optical intensity (the correlation coefficient is between 0.5 and 0.9). Subsequent investigation shows this correlation to hold independent of the presence or absence of dura mater on the samples. Poor correlation is observed between acoustic amplitude and optical intensity (the correlation coefficient is between 0.1 and 0.7).

Transcranial Assessment and Visualization of Acoustic Cavitation: Modeling and Experimental Validation.

The interaction of ultrasonically-controlled microbubble oscillations with tissues and biological media has been shown to induce a wide range of bioeffects that may have significant impact on therapy and diagnosis of brain diseases and disorders. However, the inherently non-linear microbubble oscillations combined with the micrometer and microsecond scales involved in these interactions and the limited methods to assess and visualize them transcranially hinder both their optimal use and translation to the clinics. To overcome these challenges, we present a framework that combines numerical simulations with multimodality imaging to assess and visualize the microbubble oscillations transcranially. In the present work, microbubble oscillations were studied with an integrated US and MR imaging guided clinical FUS system. A high-resolution brain CT scan was also co-registered to the US and MR images and the derived acoustic properties were used as inputs to two- and three-dimensional Finite Difference Time Domain simulations that matched the experimental conditions and geometry. Synthetic point sources by either a Gaussian function or the output of a microbubble dynamics model were numerically excited and propagated through the skull towards a virtual US imaging array. Using passive acoustic mapping (PAM) that was refined to incorporate variable speed of sound, we were able to correct the aberrations introduced by the skull and substantially improve the PAM resolution. The good agreement between the simulations incorporating microbubble emissions and experimentally-determined PAMs suggest that this integrated approach can provide a clinically-relevant framework and more control over this nonlinear and dynamic process.

Multi-planar dynamic contrast-enhanced ultrasound assessment of blood flow in a rabbit model of testicular torsion.

To assess correlation between multi-planar, dynamic contrast-enhanced ultrasound (US) blood flow measurements and radiolabeled microsphere blood flow measurements, five groups of six rabbits underwent unilateral testicular torsion of 0°, 180°, 360°, 540° or 720°. Five US measurements per testis (three transverse/two longitudinal) were obtained pre-operatively and immediately and 4 and 8 h post-operatively using linear transducers (7-4 MHz/center frequency 4.5 MHz/10 rabbits; 9-3 MHz/center frequency 5.5 MHz/20 rabbits). Björck's linear least-squares method fit the rise phase of mean pixel intensity over a 7-s period for each time curve. Slope of fit and intervention/control US pixel intensity ratios were calculated. Means of transverse, longitudinal and combined transverse/longitudinal US ratios as a function of torsion degree were compared with radiolabeled microsphere ratios using Pearson's correlation coefficient, ρ. There was high correlation between the two sets of ratios (ρ ≥ 0.88, p ≤ 0.05), except for the transverse US ratio in the immediate post-operative period (ρ = 0.79, p = 0.11). These results hold promise for future clinical applications.

A projection-based approach to diffraction tomography on curved boundaries.

An approach to diffraction tomography is investigated for two-dimensional image reconstruction of objects surrounded by an arbitrarily-shaped curve of sources and receivers. Based on the integral theorem of Helmholtz and Kirchhoff, the approach relies upon a valid choice of the Green's functions for selected conditions along the (possibly-irregular) boundary. This allows field projections from the receivers to an arbitrary external location. When performed over all source locations, it will be shown that the field caused by a hypothetical source at this external location is also known along the boundary. This field can then be projected to new external points that may serve as a virtual receiver. Under such a reformation, data may be put in a form suitable for image construction by synthetic aperture methods. Foundations of the approach are shown, followed by a mapping technique optimized for the approach. Examples formed from synthetic data are provided.

A computerized tomography system for transcranial ultrasound imaging.

Hardware for tomographic imaging presents both challenge and opportunity for simplification when compared with traditional pulse-echo imaging systems. Specifically, point diffraction tomography does not require simultaneous powering of elements, in theory allowing just a single transmit channel and a single receive channel to be coupled with a switching or multiplexing network. In our ongoing work on transcranial imaging, we have developed a 512-channel system designed to transmit and/or receive a high voltage signal from/to arbitrary elements of an imaging array. The overall design follows a hierarchy of modules including a software interface, microcontroller, pulse generator, pulse amplifier, high-voltage power converter, switching mother board, switching daughter board, receiver amplifier, analog-to-digital converter, peak detector, memory, and USB communication. Two pulse amplifiers are included, each capable of producing up to 400Vpp via power MOSFETS. Switching is based around mechanical relays that allow passage of 200V, while still achieving switching times of under 2ms, with an operating frequency ranging from below 100kHz to 10MHz. The system is demonstrated through ex vivo human skulls using 1MHz transducers. The overall system design is applicable to planned human studies in transcranial image acquisition, and may have additional tomographic applications for other materials necessitating a high signal output.

Two-dimensional image reconstruction with spectrally-randomized ultrasound signals.

An ultrasound imaging method using unfocused frequency-randomized transmissions and image reconstruction from data received by a single element is experimentally demonstrated. The elements of an ultrasound imaging array are randomly assigned different frequencies and driven by a multicycle sinusoidal burst. The resulting acoustic field is spectrally unique and target localization is possible based on the a priori knowledge of this field. A 64-element phased array driven by arbitrary waveform generators is used in the experiments. Transmission frequencies range from 2.00 to 2.64 MHz with 10 kHz resolution. One element of the array is reserved for receiving backscattered signals and an image is reconstructed from the signals received by this single element. Reconstruction is based on cross-correlation of the received data with transmitted bursts to obtain radial elliptical projections. Multiple projections are obtained from single received data, which are back-projected to obtain an image. Successful target localization is made possible through multiple frequency-randomized acquisitions. The performance of the method is measured using images of a single point target. These images are quantified and analyzed in terms of their point spread function (PSF) and SNR. Optimum imaging parameters, such as the number of acquisitions, transmit burst length, and number of possible receivers, are obtained through further analysis of SNR. Images obtained with the frequency-randomized transmission method compared well with the performance measurements of a typical B-mode acquisition. It is demonstrated that the frequency-randomized method provides images superior to B-mode images in terms of PSF. The two-point discrimination threshold is measured to be 2 mm in the lateral and azimuth directions.

Application of the split-step Padé approach to nonlinear field predictions.

We herein propose a new theoretical approach for analyzing the nonlinear propagation of directive sound beams emitted from a planar piston source with a circular aperture. The proposed approach relies on the split-step Padé approximation, which is an efficient method for obtaining wide-angle one-way wave equations, especially in underwater acoustics. Despite including only two Padé terms in the expansion, the theory was applicable to a beam angle of up to ±40° relative to the main propagation direction, the angle of which is approximately twice that of the Khokhlov-Zabolotskaya-Kuznetsov equation, which is based on parabolic approximation. In order to demonstrate the effectiveness of the newly proposed theoretical approach, we performed an experiment using an airborne ultrasonic emitter with a circular aperture of 7.5cm in radius. We drove the emitter powerfully at a 36-kHz and 40-kHz bi-frequency signal and measured the beam patterns of the primary and secondary waves, such as parametric sounds within wide propagation angles. Excellent agreement between measured data and the corresponding numerical simulations supports the validity of the proposed model equations and the computational methods for their numerical solutions.

The feasibility of pulse compression by nonlinear effective bandwidth extension.

Chirp-encoded excitation has been utilized for increased signal-to-noise ratio (SNR) in both linear and harmonic imaging. In either case, it is necessary to isolate the relevant frequency band to avoid artifacts. In contrast, the present study isolates and then combines the fundamental and the higher harmonics, treating them as a single, extended bandwidth. Pulse-inverted sum and difference signals are first used to isolate even and odd harmonics. Matched filters specific to the source geometry and the transmit signal are then separately applied to each harmonic band. Verification experiments are performed using up to the third harmonic resulting from an underwater chirp excitation. Analysis of signal peaks after scattering from a series of steel and nylon wires indicates increased compression using the extended bandwidth, as compared to well-established methods for fundamental and second harmonic chirp compression. Using third harmonic bands, a mean pulse width of 56% relative to fundamental compression and 48% relative to second harmonic compression was observed. Further optimization of the compression by altering the transmission indicated 17% additional reduction in the pulse width and a 47% increase in peak-to-sidelobe ratio. Overall, results establish the feasibility of extended bandwidth signal compression for simultaneously increasing SNR and signal resolution.

A wireless batteryless deep-seated implantable ultrasonic pulser-receiver powered by magnetic coupling.

This study tests a deep-seated implantable ultrasonic pulser-receiver, powered wirelessly by magnetic coupling. A 30-cm energy-transmitting coil was designed to wrap around the body, and was driven by a current of 1.2 A rms at a frequency of 5.7 MHz to generate a magnetic field. A 2-cm receiving coil was positioned at the center of the primary coil for receiving the magnetic energy and powering the implantable device. A capacitor-diode voltage multiplier in the implantable circuit was used to step-up the receiving coil voltage from 12.5 to 50 V to operate an ultrasonic pulser. FEA magnetic field simulations, bench-top, and ex vivo rabbit measurements showed that the magnetic energy absorption in body tissue is negligible and that the magnetic coupling is not sensitive to receiving coil placement. The receiving coil and the power conditioning circuits in the implantable device do not contain ferromagnetic material, so a magnetic-resonance-compatible device can be achieved. A 5-MHz ultrasound transducer was used to test the implantable circuit, operating in pulse-echo mode. The received echo was amplified, envelope-detected, frequency-modulated, and transmitted out of the rabbit body by a radio wave. The modulated echo envelope signal was received by an external receiver located about 10 cm away from the primary coil. The study concludes that operation of a batteryless and wireless deep-seated implantable ultrasonic pulser-receiver powered by coplanar magnetic coupling is feasible.

Spatial backward planar projection in absorbing media possessing an arbitrary dispersion relation.

Planar projection methods have been shown to rapidly relate fields between two planes. Such an approach is particularly useful for characterizing transducers, since only a single plane needs to be measured in order to characterize an entire field. The present work considers the same approach in the presence of an arbitrary dispersion relation. Unlike traditional methods that use Fourier solutions of the time-domain wave equation, the approach starts from a frequency-domain Helmholtz equation for waves in a dispersive medium. It is shown that a transfer function similar to that derived from time domain equations can be utilized. Both the forward- and backward-projection behaviors are examined and it is demonstrated that the approach is invariant to propagation direction.

Standing-wave suppression for transcranial ultrasound by random modulation.

Low-frequency transcranial ultrasound (<<1 MHz) is being investigated for a number of brain therapies, including stroke, tumor ablation, and localized opening of the blood-brain barrier. However, lower frequencies have been associated with the production of undesired standing waves and cavitation in the brain. Presently, we examine an approach to suppress standing waves during continuous-wave (CW) transcranial application. The investigation uses a small randomization in the frequency content of the signal for suppressing standing waves. The approach is studied in an ex-vivo human skull and a plastic-walled chamber, representing idealized conditions. The approach is compared to single-frequency CW operation as well as to a swept-frequency input. Acoustic field scans demonstrate that the swept-frequency method can suppress standing waves in the plastic chamber and skull by 3.4 and 1.6 times, respectively, compared to single-frequency CW excitation. With random modulation, standing waves were reduced by 5.6 and 2 times, respectively, in the plastic chamber and skull. It is expected that the process may play a critical role in providing a safer application of the ultrasound field in the brain and may have application in other areas where standing waves may be created.

Accelerated focused ultrasound imaging.

One of the most basic trade-offs in ultrasound imaging involves frame rate, depth, and number of lines. Achieving good spatial resolution and coverage requires a large number of lines, leading to decreases in frame rate. An even more serious imaging challenge occurs with imaging modes involving spatial compounding and 3-D/4-D imaging, which are severely limited by the slow speed of sound in tissue. The present work can overcome these traditional limitations, making ultrasound imaging many-fold faster. By emitting several beams at once, and by separating the resulting overlapped signals through spatial and temporal processing, spatial resolution and/or coverage can be increased by many-fold while leaving frame rates unaffected. The proposed approach can also be extended to imaging strategies that do not involve transmit beamforming, such as synthetic aperture imaging. Simulated and experimental results are presented where imaging speed is improved by up to 32-fold, with little impact on image quality. Object complexity has little impact on the method's performance, and data from biological systems can readily be handled. The present work may open the door to novel multiplexed and/or multidimensional protocols considered impractical today.

Acoustic standing wave suppression using randomized phase-shift-keying excitations.

Recent papers have demonstrated that acoustic standing waves can be inhibited by frequency-modulated spread-spectrum excitation. An alternative method is studied here that is designed to be more practical for implementation in phased arrays. The method operates using phase-shift-keying (PSK), which introduces phase shifts into the driving signal to break wave symmetry. Sequential and random binary-PSK (BPSK) and quadrature-PSK (QPSK) excitations are studied in water, using a carrier frequency of 250 kHz and a time segment of 10 cycles. The resulting acoustic field is measured with a transducer inside a plastic-walled chamber and compared with continuous wave excitation. Results indicate that both the random BPSK and QPSK methods can reduce time-averaged spatial intensity variation caused by standing waves by approximately six times.

Ultrasound phase contrast thermal imaging with reflex transmission imaging methods in tissue phantoms.

Thermal imaging measurements using ultrasound phase contrast have been performed in tissue phantoms heated with a focused ultrasound source. Back projection and reflex transmission imaging principles were used to detect sound speed-induced changes in the phase caused by an increase in the temperature. The temperature was determined from an empirical relationship for the temperature dependence on sound speed. The phase contrast was determined from changes in the sound field measured with a hydrophone scan conducted before and during applied heating. The lengthy scanning routine used to mimic a large two-dimensional array required a steady-state temperature distribution within the phantom. The temperature distribution in the phantom was validated with magnetic resonance (MR) thermal imaging measurements. The peak temperature was found to agree within 1 degrees C with MR, and good agreement was found between the temperature profiles. The spatial resolution was 0.3x0.3x0.3mm, comparing favorably with the 0.625x0.625x1.5-mm MR spatial resolution.

A harmonic cancellation technique for an ultrasound transducer excited by a switched-mode power converter.

The aim of this study is to evaluate the feasibility of using harmonic cancellation for a therapeutic ultrasound transducer excited by a switched-mode power converter without an additional output filter. A switching waveform without the third harmonic was created by cascading two switched-mode power inverter modules at which their output waveforms were pi/3 phase shifted from each other. A PSPICE simulation model for the power converter output stage was developed. The simulated results were in good agreement with the measurement. The waveform and harmonic contents of the acoustic pressure generated by a 1-MHz, self-focused piezoelectric transducer with and without harmonic cancellation have been evaluated. Measured results indicated that the acoustic third harmonicto- fundamental ratio at the focus was small (-48 dB) with harmonic cancellation, compared to that without harmonic cancellation (-20 dB). The measured acoustic levels of the fifth harmonic for both cases with and without harmonic cancellation also were small (-46 dB) compared to the fundamental. This study shows that it is viable to drive a piezoelectric ultrasound transducer using a switched-mode power converter without the requirement of an additional output filter in many high-intensity focused ultrasound (HIFU) applications.

A computer-controlled ultrasound pulser-receiver system for transskull fluid detection using a shear wave transmission technique.

The purpose of this study was to evaluate the performance of a computer-controlled ultrasound pulser-receiver system incorporating a shear mode technique for transskull fluid detection. The presence of fluid in the sinuses of an ex vivo human skull was examined using a pulse-echo method by transmitting an ultrasound beam through the maxilla bone toward the back wall on the other side of the sinus cavity. The pulser was programmed to generate bipolar pulse trains with 5 cycles at a frequency of 1 MHz, repetition frequency of about 20 Hz, and amplitude of 100 V to drive a 1-MHz piezoelectric transducer. Shear and longitudinal waves in the maxilla bone were produced by adjusting the bone surface incident angle to 45 degrees and 0 degrees, respectively. Computer tomography (CT) scans of the skull were performed to verify the ultrasound experiment. Using the shear mode technique, the echo waveform clearly distinguishes the presence of fluid, and the estimated distance of the ultrasound traveled in the sinus is consistent with the measurement from the CT images. Contrarily, using the longitudinal mode, no detectable back wall echo was observed under the same conditions. As a conclusion, this study demonstrated that the proposed pulser-receiver system with the shear mode technique is promising for transskull fluid detecting, such as mucus in a sinus.

Two-dimensional ultrasound detection with unfocused frequency-randomized signals.

A method is described for detecting scattering in two-dimensions using an unfocused ultrasound field created from a continuously driven source array. The frequency of each element on the array is unique, resulting in a field that is highly variant as a function of both time and position. The scattered signal is then received by a single receiving line. The method, as currently written, is valid under the first order Born approximation. To demonstrate the approach, a series of simulations within the frequency range of 0.10-1.25 MHz are performed and compared with a simulated B-Scan in the same frequency range. The method is found to be superior in resolving closely spaced objects, discerning 1.4 mm separation in the radial and 0.5-mm separation in the axial direction. The method was also better able to determine object size, resolving scatters less than 10% of wavelength associated with the center frequency.