Resolving Ultrasound Contrast Microbubbles Using Minimum Variance Beamforming.

Minimum Variance (MV) beamforming is known to improve the lateral resolution of ultrasound images and enhance the separation of isolated point scatterers. This paper aims to evaluate the adaptive beamformer's performance with flowing microbubbles (MBs) which are relevant to super-resolution ultrasound imaging. Simulations using point scatterer data from single emissions were complemented by an experimental investigation performed using a capillary tube phantom and the Synthetic Aperture Real-time Ultrasound System (SARUS). The MV performance was assessed by the minimum distance that allows the display of two scatterers positioned side-by-side, the lateral Full-Width-at-Half-Maximum (FWHM), and the Peak-Sidelobe-Level (PSL). In the tube, scatterer responses separated by down to [Formula: see text] (or 1.05λ ) were distinguished by the MV method, while the standard Delay-And-Sum (DAS) beamformers were unable to achieve such separation. Up to ninefold FWHM decrease was also measured in favor of the MV beamformer for individual echoes from MBs. The lateral distance between two scatterers impacted on their FWHM value, and additional differences in the scatterers' axial or out-of-plane position also impacted on their size and appearance. The simulation and experimental results were in agreement in terms of lateral resolution. The point scatterer study showed that the proposed MV imaging scheme provided clear resolution benefits compared to DAS. Current super-resolution methods mainly depend on DAS beamformers. Instead, the use of the MV method may provide a larger number of detected, and potentially better localized, MB scatterers.

Influence of temperature, needle gauge and injection rate on the size distribution, concentration and acoustic responses of ultrasound contrast agents at high frequency.

This paper investigated the influence of needle gauge (19G and 27G), injection rate (0.85ml·min(-1), 3ml·min(-1)) and temperature (room temperature (RT) and body temperature (BT)) on the mean diameter, concentration, acoustic attenuation, contrast to tissue ratio (CTR) and normalised subharmonic intensity (NSI) of three ultrasound contrast agents (UCAs): Definity, SonoVue and MicroMarker (untargeted). A broadband substitution technique was used to acquire the acoustic properties over the frequency range 17-31MHz with a preclinical ultrasound scanner Vevo770 (Visualsonics, Canada). Significant differences (P<0.001-P<0.05) between typical in vitro setting (19G needle, 3ml·min(-1) at RT) and typical in vivo setting (27G needle, 0.85ml·min(-1) at BT) were found for SonoVue and MicroMarker. Moreover we found that the mean volume-based diameter and concentration of both SonoVue and Definity reduced significantly when changing from typical in vitro to in vivo experimental set-ups, while those for MicroMarker did not significantly change. From our limited measurements of Definity, we found no significant change in attenuation, CTR and NSI with needle gauge. For SonoVue, all the measured acoustic properties (attenuation, CTR and NSI) reduced significantly when changing from typical in vitro to in vivo experimental conditions, while for MicroMarker, only the NSI reduced, with attenuation and CTR increasing significantly. These differences suggest that changes in physical compression and temperature are likely to alter the shell structure of the UCAs resulting in measureable and significant changes in the physical and high frequency acoustical properties of the contrast agents under typical in vitro and preclinical in vivo experimental conditions.

Laser Doppler anemometry measurements of the shear stresses on ultrasonic contrast agent microbubbles attached to agar.

Ultrasonic contrast agents are currently being developed to target and bind to specific areas of interest such as atheromous plaque. A microbubble has been developed in-house which can be targeted to attach to specific cell-lines. To assess the feasibility of using the microbubble in vivo, the shear stresses which the bound microbubbles can withstand need to be known. A flow chamber was developed for use with intravascular ultrasound (IVUS) and laser Doppler anemometry (LDA). Biotin was incorporated into the microbubble shells and streptavidin was used to attach them to agar. IVUS at 40 MHz was then used to image the attached microbubbles under steady flow at a range of flow rates from 75 to 480 mL min(-1) through a flow area of 9 mm(2). LDA was employed to find high resolution velocity profiles of the flow in the chamber at a selection of these flow rates and the shear stresses on the bubbles were calculated. The bubbles were found to remain attached to the agar for shear stresses of up to 3.4 Pa. This compares with mean physiological arterial shear stresses of less than 1.5 Pa for pulsatile flow.

The polydisperse acoustic signature of rigid microbubbles.

Microbubbles are used in medical ultrasound imaging as contrast agents to image the vascular bed under the mode of Ultrasound Contrast Imaging (UCI). The microbubble shell determines the acoustic response and hence the signal that is utilized to form the images in UCI. Single microbubble signals from BiSphere™ (POINT Biomedical, San Carlos, CA, USA) microbubbles were captured using a clinical ultrasound system. Three main typical responses of microbubbles were identified, a) full duration echo, b) echo with duration shorter than the incident pulse and c) echo that in part resembles that in (b) and in addition prior to that another short duration initial lower amplitude signal. These data corroborate that the shell structural and nanomechanical property provide the different responses at different microbubble sizes. These different signals present an opportunity for tracking the movement of well differentiated single microbubbles particularly with novel super-resolution imaging methods that require sparse microbubble populations.

In vitro acoustic characterization of three phospholipid ultrasound contrast agents from 12 to 43 MHz.

The acoustic properties of two clinical (Definity, Lantheus Medical Imaging, North Billerica, MA, USA; SonoVue, Bracco S.P.A., Milan, Italy) and one pre-clinical (MicroMarker, untargeted, Bracco, Geneva, Switzerland; VisualSonics, Toronto, ON, Canada) ultrasound contrast agent were characterized using a broadband substitution technique over the ultrasound frequency range 12-43 MHz at 20 ± 1°C. At the same number concentration, the acoustic attenuation and contrast-to-tissue ratio of the three native ultrasound contrast agents are comparable at frequencies below 30 MHz, though their size distributions and encapsulated gases and shells differ. At frequencies above 30 MHz, native MicroMarker has higher attenuation values and contrast-to-tissue ratios than native Definity and SonoVue. Decantation was found to be an effective method to alter the size distribution and concentration of native clinical microbubble populations, enabling further contrast enhancement for specific pre-clinical applications.

A setup for the assessment of the effect of tubular confinement on the acoustic response of microbubbles.

Ultrasound contrast agents are gas filled microbubbles which produced enhanced echoes in ultrasound imaging thus allowing the acquisition of detailed information on the path of blood. It is theoretically known that the size of a vessel affects the behavior of a microbubble, which could potentially be used to discriminate different sized vessels. This information would be useful in the monitoring of neovascularization in tumor growth or treatment. However, currently it is not possible to identify the vessel diameter by any means of signal processing of microbubble echoes. In order to assess microbubble behavior when confined in tubes we compared the acoustic backscatter from biSphere™ microbubbles both free in water and flowing in 200 µm diameter tubes that are similar in size to arterioles. Experimental systems that allow the interrogation of individual microbubbles were designed and modified to allow investigation of both free microbubbles and those in tubes. Unprocessed single microbubble RF data were collected, allowing the calculation of both the fundamental and second harmonic components of the backscattered signal. Microbubbles confined in tubes had lower amplitude response compared to unconfined microbubbles. On consecutive insonations of the same microbubble, free microbubbles produced echoes above noise more often than confined microbubbles. This setup may be used to investigate microbubble behavior in a range of smaller tubes with diameters similar to capillaries thus enabling signal processing design for vessel differentiation.

The speed of sound and attenuation of an IEC agar-based tissue-mimicking material for high frequency ultrasound applications.

This study characterized the acoustic properties of an International Electromechanical Commission (IEC) agar-based tissue mimicking material (TMM) at ultrasound frequencies in the range 10-47 MHz. A broadband reflection substitution technique was employed using two independent systems at 21°C ± 1°C. Using a commercially available preclinical ultrasound scanner and a scanning acoustic macroscope, the measured speeds of sound were 1547.4 ± 1.4 m∙s(-1) and 1548.0 ± 6.1 m∙s(-1), respectively, and were approximately constant over the frequency range. The measured attenuation (dB∙cm(-1)) was found to vary with frequency f (MHz) as 0.40f + 0.0076f(2). Using this polynomial equation and extrapolating to lower frequencies give values comparable to those published at lower frequencies and can estimate the attenuation of this TMM in the frequency range up to 47 MHz. This characterisation enhances understanding in the use of this TMM as a tissue equivalent material for high frequency ultrasound applications.