Newly Designed 3D-Printed Sonication Test Cell Optimized for In Vitro Sonication Experiments.

OBJECTIVE: Precise control over the ultrasound field parameters experienced by biological samples during sonication experiments in vitro may be quite challenging. The main goal of this work was to outline an approach to construction of sonication test cells that would minimize the interaction between the test cells and ultrasound. METHODS: Optimal dimensions of the test cell were determined through measurements conducted in a water sonication tank using 3D-printed test objects. The offset of local acoustic intensity variability inside the sonication test cell was set to value of ±50% of the reference value (i.e., local acoustic intensity measured at last axial maximum in the free-field condition). The cytotoxicity of several materials used for 3D printing was determined using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay. RESULTS: The sonication test cells were 3D printed from polylactic acid material, which was not toxic to the cells. Silicone membrane HT-6240, which was used to construct the bottom of the test cell, was found to reduce ultrasound energy minimally. Final ultrasound profiles inside the sonication test cells indicated the desired variability of local acoustic intensity. The cell viability in our sonication test cell was comparable to that of commercial culture plates with bottoms constructed with silicone membrane. CONCLUSION: An approach to construction of sonication test cells minimizing the interaction of the test cell and ultrasound has been outlined.

Far field during sonication experiments in vitro - Is it really far enough?

In many in vitro experiments studying ultrasound bioeffects the sonicated samples are placed to far field with intention of exposing them to as uniform ultrasound field as possible. The main aim of this work is to assess whether the sonicated samples really experience what they are believed to. Also we would like to suggest basic rules for construction of sonication vessels. We used 3.5 MHz and 7 MHz ultrasound transducers for measurements. We measured ultrasound field inside and behind common culture plates and special 3D printed plates placed to last axial maximum in water sonication tank with use of a needle hydrophone. Our results show that even though the sonication vessels with sonicated samples are placed into far field, the sonicated samples are actually exposed to some kind of a near field pattern which develops due to the interaction between ultrasound and well of culture plate. The variability of local acoustic intensity can reach up to several hundreds of percent. Our results are also supported by theoretical calculation and software for simulation of ultrasound fields. Even though the sonicated samples may have actually been exposed to some kind of near field pattern in many past studies, the whole phenomenon of creation of near field pattern can be controlled to some extent for future studies. Thus, we suggest that the sonication vessel should always be designed for particular ultrasound transducer.

Therapeutic ultrasound experiments in vitro: Review of factors influencing outcomes and reproducibility.

Current in vitro sonication experiments show immense variability in experimental set-ups and methods used. As a result, there is uncertainty in the ultrasound field parameters experienced by sonicated samples, poor reproducibility of these experiments and thus reduced scientific value of the results obtained. The scope of this narrative review is to briefly describe mechanisms of action of ultrasound, list the most frequently used experimental set-ups and focus on a description of factors influencing the outcomes and reproducibility of these experiments. The factors assessed include: proper reporting of ultrasound exposure parameters, experimental geometry, coupling medium quality, influence of culture vessels, formation of standing waves, motion/rotation of the sonicated sample and the characteristics of the sample itself. In the discussion we describe pros and cons of particular exposure geometries and factors, and make a few recommendations as to how to increase the reproducibility and validity of the experiments performed.

Evaluation of the Effect of Tissue Compression on the Results of Shear Wave Elastography Measurements.

Shear wave imaging is considered to be more precise and less operator dependent when compared with strain imaging. It enables quantitative and reproducible data (Young's modulus of the imaged tissue). However, results of shear wave imaging can be affected by a variety of different factors. The aim of this study is to evaluate the effect of the pressure applied by the ultrasound probe during examination on the measured values of Young's modulus. The effect of the tissue compression on the results of the real-time shear wave elastography was evaluated via the gelatine phantom measurements, via the ex vivo experiments with pig liver, and via the in vivo measurements of the thyroid gland stiffness on healthy volunteers. The results of our measurements confirmed that the measured value of Young's modulus increases with the increasing pressure applied on the imaged object. The highest increase was observed during the ex vivo experiments (400%), and the lowest increase was detected in the case of the phantom measurements (8%). A two- to threefold increase in Young's modulus was observed between the minimum and maximum pressure in the case of the in vivo elastography measurements of thyroid gland. The Veronda-Westman theoretical model was used for the description of the tissue nonlinearity. We conclude that tissue compression by the force exerted on the probe can significantly affect the results of the real-time shear wave elastography measurements. Minimum pressure should be used when measuring the absolute value of Young's modulus of superficial organs.

The effect of dead elements on the accuracy of Doppler ultrasound measurements.

The objective of this study is to investigate the effect of multiple dead elements in an ultrasound probe on the accuracy of Doppler ultrasound measurements. For this work, we used a specially designed ultrasound imaging system, the Ultrasonix Sonix RP, that provides the user with the ability to disable selected elements in the probe. Using fully functional convex, linear, and phased array probes, we established a performance baseline by measuring the parameters of a laminar parabolic flow profile. These same parameters were then measured using probes with 1 to 10 disabled elements. The acquired velocity spectra from the functional probes and the probes with disabled elements were then analyzed to determine the overall Doppler power, maximum flow velocity, and average flow velocity. Color Flow Doppler images were also evaluated in a similar manner. The analysis of the Doppler spectra indicates that the overall Doppler power as well as the detected maximum and average velocities decrease with the increasing number of disabled elements. With multiple disabled elements, decreases in the detected maximum and average velocities greater than 20% were recorded. Similar results were also observed with Color Flow Doppler measurements. Our results confirmed that the degradation of the ultrasound probe through the loss of viable elements will negatively affect the quality of the Doppler-derived diagnostic information. We conclude that the results of Doppler measurements cannot be considered accurate or reliable if there are four or more contiguous dead elements in any given probe.

A contribution to sonograph image quality estimation using point spread function.

In medical sonography, sonograph image quality is an essential aspect for the safety of both patient and doctor. Its evaluation therefore requires an accurate and objective method for measurement. In this regard, a number of methods are in current use. Most of these are based on tissue mimicking phantom imaging. In contrast, we have used another principle based on Point Spread Function (PSF) analysis which is a product of the measuring system we have developed. In this case, the measured sonograph scans a small metallic ball target that moves in a water bath on a specified trajectory. The Region Of Interest (ROI) of the sonogram containing the ball target picture is digitised and the amplitude of the pixels analysed. The result is the PSF from which we calculate the lateral resolution (LR). For this purpose, we use our own original software. Using this method, we have to date been able to plot LR characteristics over the scanning plane. The method allows us to differentiate separate scanning lines and even multiple focal areas for dynamic focussing systems. It can detect malfunctions in dynamic focussing, size of aperture, time gain compensation function and/or transducer element failure. The procedure itself is not as easy or as fast to use as tissue mimicking phantoms or 3D signal to noise ratio evaluation, but it provides accurate and objective numeric parameters corresponding to the quality of image at any specified point over the whole scanning area. It is also a very powerful tool when used in combination with the other methods mentioned above.