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This work focuses on the use of ultrasound imaging to evaluate the cell concentration of dilute leukocyte suspensions in the range of 10-3000 cells/µL. First, numerical simulations were used to study the influence of the size dispersion and the leukocyte type on the performance of the concentration estimation algorithms, which were developed in previous works assuming single-sized scatterers. From this analysis, corrections to the mentioned algorithms were proposed and then the performance of these corrections was evaluated from experiments. For this, ultrasound images were captured from suspensions of lymphocytes, granulocytes, and their mixtures. These images were obtained using a 20 MHz single-channel scanning system. Results confirmed that concentration estimates provided by conventional algorithms were affected by the size dispersion of cells, leading to a remarkable underestimation of results. The proposed correction to compensate for cell size dispersion obtained from simulations improved the concentration estimation of these algorithms, for the cell suspensions tested, approaching the results to the reference optical characterization. Moreover, it was shown that these models provided a total leukocyte concentration from the ultrasound images which was independent of the relative populations of different white blood cell types.

This work deals with the development of a methodology to evaluate the concentration in cell or particle suspensions from ultrasound images. The novelty of the method is based on two goals: first, it should be valid when the energy reaching the scatterers is unknown and cannot be measured or calibrated. In addition, it should be robust against echo overlap which may occur due to high scatterer concentration. Both characteristics are especially valuable in quantitative ultrasound analysis in the clinical context. In this regard, the present work considers the ability of envelope statistics models to characterize ultrasound images. Envelope statistical analysis are based on the examination of the physical properties of a medium through the study of the statistical distribution of the backscattered signal envelop. A review of the statistical distributions typically used to characterize scattering mediums was conducted. The main parameters of the distribution were estimated from simulations of signals backscattered by particle suspensions. Then, the ability of these parameters to characterize the suspension concentration was analyzed and the µ parameter from the Homodyned-K distribution resulted as the most suitable parameter for the task. Simulations were also used to study the impact of noise, signal amplitude variability and dispersion of particle sizes on the estimation method. The efficiency of the algorithm on experimental measurements was also evaluated. To this end, two sets of ultrasound images were obtained from suspensions of 7 µm and 12 µm polystyrene particles in water, using a 20 MHz focused transducer. The methodology proved to be efficient to quantify the concentration of particle suspensions in the range between 5 and 3000 particles/µl, achieving similar results for both particle sizes and for different signal-to-noise ratios., (Copyright © 2021 Elsevier B.V. All rights reserved.)

A methodology for the assessment of cell concentration, in the range 5-100 cells/ [Formula: see text], suitable for in vivo analysis of serous body fluids is presented in this work. This methodology is based on the quantitative analysis of ultrasound images obtained from cell suspensions and considers applicability criteria, such as short analysis times, moderate frequency, and absolute concentration estimation, all necessary to deal with the variability of tissues among different patients. Numerical simulations provided the framework to analyze the impact of echo overlapping and the polydispersion of scatterer sizes on the cell concentration estimation. The cell concentration range that can be analyzed as a function of the transducer and emitted waveform used was also discussed. Experiments were conducted to evaluate the performance of the method using 7- [Formula: see text] and 12- [Formula: see text] polystyrene particles in water suspensions in the 5-100 particles/ [Formula: see text] range. A single scanning focused transducer working at a central frequency of 20 MHz was used to obtain ultrasound images. The method proposed to estimate the concentration proved to be robust for different particle sizes and variations of gain acquisition settings. The effect of tissues placed in the ultrasound path between the probe and the sample was also investigated using 3-mm-thick tissue mimics. Under this situation, the algorithm was robust for the concentration analysis of 12 [Formula: see text] particle suspensions, yet significant deviations were obtained for the smallest particles.

Ultrasonic phase velocity spectroscopy is a very sensitive technique used in the measurement of material properties. In a phase velocity calculation, ambiguities can arise in the spectral phases, in the form of integer multiples of 2π rad, which, if not corrected, results in large errors. In this work, we propose a method for determining these ambiguities, more specifically, the number of 2π rad phase jumps, using the Kramers-Kronig relations, for samples exhibiting a frequency power-law attenuation coefficient. The method is based on a first estimate of the phase velocity from group velocity and attenuation coefficient that are not affected by phase jumps. This estimated phase velocity is used to obtain the number of 2π rad phase jumps, which in turn is used to calculate the corrected phase velocity. The method was tested with samples of liquids with a frequency power-law attenuation coefficient (exponent y varying from 1.5 to 2) and a solid [polymethyl methacrylate (PMMA)] with y  ∼  1 , and velocity dispersions ranging from 0 to 34 (cm/s)/MHz.

This work presents an affordable and easily customizable methodology for phantom manufacturing, which can be used to mimic different anatomic organs and structures. This methodology is based on the use of polyvinyl alcohol-based cryogels as a physical substitute for biologic soft tissues and of 3-D printed polymers for hard tissues, moulding and supporting elements. Thin and durable soft-tissue mimicking layers and multilayer arrangements can be obtained using these materials. Special attention was paid to the acoustic properties (sound speed, attenuation coefficient and mechanical impedance) of the materials developed to simulate soft tissues. These properties were characterized as a function of the additives concentration (propylene-glycol and alumina particles). The polyvinyl alcohol formulation proposed in this work is stable over several freeze-thaw cycles, allowing the manufacturing of multilayer materials with controlled properties. The manufacturing methodology presented was applied to the development of a phantom for high-frequency cranial ultrasonography in infants. This phantom was able to reproduce the main characteristics of the ultrasound images obtained in neonates through the anterior fontanel, down to 8-mm depth., (Copyright © 2019 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

This work analyzes some key aspects of the behavior of sensors based on piezoelectric Thickness Shear Mode (TSM) resonators to study and monitor microbial biofilms. The operation of these sensors is based on the analysis of their resonance properties (both resonance frequency and dissipation factor) that vary in contact with the analyzed sample. This work shows that different variations during the microorganism growth can be detected by the sensors and highlights which of these changes are indicative of biofilm formation. TSM sensors have been used to monitor in real time the development of Staphylococcus epidermidis and Escherichia coli biofilms, formed on the gold electrode of the quartz crystal resonators, without any coating. Strains with different ability to produce biofilm have been tested. It was shown that, once a first homogeneous adhesion of bacteria was produced on the substrate, the biofilm can be considered as a semi-infinite layer and the quartz sensor reflects only the viscoelastic properties of the region immediately adjacent to the resonator, not being sensitive to upper layers of the biofilm. The experiments allow the microrheological evaluation of the complex shear modulus ( G * = G ' + jG ″) of the biofilm at 5 MHz and at 15 MHz, showing that the characteristic parameter that indicates the adhesion of a biofilm for the case of S. epidermidis and E. coli , is an increase in the resonance frequency shift of the quartz crystal sensor, which is connected with an increase of the real shear modulus, related to the elasticity or stiffness of the layer. In addition both the real and the imaginary shear modulus are frequency dependent at these high frequencies in biofilms., Competing Interests: The authors declare no conflict of interest.

Accurate measurement of very low cerebrospinal fluid (CSF) white blood cell (WBC) concentration is key to the diagnosis of bacterial meningitis, lethal if not promptly treated. Here we show that high frequency ultrasound (HFUS) can detect CSF WBC in vitro in concentrations relevant to meningitis diagnosis with a much finer precision than gold standard manual counting in a Fuchs-Rosenthal chamber. WBC concentrations in a mock CSF model, in the range 0-50 WBC/μL, have been tested and compared to gold standard ground truth. In this range, excellent agreement (Cohen's kappa [κ] = 0.78-90) (Cohen 1960) was observed between HFUS and the gold standard method. The presented experimental set-up allowed us to detect WBC concentrations as low as 2 cells/μL. HFUS shows promise as a low-cost, reliable and automated technology to measure very low CSF WBC concentrations for the diagnosis of early meningitis., (Copyright © 2016 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

This work proposes the use of an ultrasound based technique to measure the concentration of yeasts in liquid suspension. This measurement was achieved by the detection and quantification of ultrasonic echoes backscattered by the cells. More specifically, the technique was applied to the detection and quantification of Saccharomyces cerevisiae. A theoretical approach was proposed to get the average density and sound speed of the yeasts, which were found to be 1116 kg/m(3) and 1679 m/s, respectively. These parameters were needed to model the waves backscattered by each single cell. A pulse-echo arrangement working around 50 MHz, being able to detect echoes from single yeasts was used to characterize experimentally yeast solutions from 10(2) to 10(7)cells/ml. The Non-negative Matrix Factorization denoising technique was applied for data analysis. This technique required a previous learning of the spectral patterns of the echoes reflected from yeasts in solution and the base noise from the liquid medium. Comparison between pulse correlation (without denoising) and theoretical and experimental pattern learning was made to select the best signal processing. A linear relation between ultrasound output and concentration was obtained with correlation coefficient R(2)=0.996 for the experimental learning. Concentrations from 10(4) to 10(7)cells/ml were detected above the base noise. These results show the viability of using the ultrasound backscattering technique to detect yeasts and measure their concentration in liquid cultures, improving the sensitivity obtained using spectrophotometric methods by one order of magnitude., (Copyright © 2015 Elsevier B.V. All rights reserved.)

This work proposes the use of quartz crystal microbalances (QCMs) as a method to analyze and characterize magnetorheological (MR) fluids. QCM devices are sensitive to changes in mass, surface interactions, and viscoelastic properties of the medium contacting its surface. These features make the QCM suitable to study MR fluids and their response to variable environmental conditions. MR fluids change their structure and viscoelastic properties under the action of an external magnetic field, this change being determined by the particle volume fraction, the magnetic field strength, and the presence of thixotropic agents among other factors. In this work, the measurement of the resonance parameters (resonance frequency and dissipation factor) of a QCM are used to analyze the behavior of MR fluids in static conditions (that is, in the absence of external mechanical stresses). The influence of sedimentation under gravity and the application of magnetic fields on the shifts of resonance frequency and dissipation factor were measured and discussed in the frame of the coupled resonance produced by particles touching the QCM surface. Furthermore, the MR-fluid/QCM system has a great potential for the study of high-frequency contact mechanics because the translational and rotational stiffness of the link between the surface and the particles can be tuned by the magnetic field.

The effect of particle volume fraction on the microstructure of magnetorheological (MR) fluids has been studied using ultrasonic techniques. When no magnetic field is applied, they behave as slurry. However, when magnetic field is applied, important features regarding the change of the microstructure have been found with the help of ultrasonic waves propagating in the direction of the magnetic field. As the volume fraction increases, a rearrangement of particles which decrease the compressibility of the system is detected; nevertheless, the material behaves as a non-consolidated material. Three different particle volume fraction regions are found identifying a critical particle volume fraction predicted in the literature. Ultrasounds are confirmed as an interesting tool to study MR fluids in static conditions., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Comment is made on an article by Temkin [J. Acoust. Soc. Am. 108(1), 126-146 (2000)]. This work illustrates the impact of particle size distribution on the dispersion and attenuation of sound waves propagating through particle suspensions. Temkin's propagation theory is extended for poly-disperse distributions of spherical particles, applicable to real suspensions. Significant changes in dispersion and attenuation appear near the particle resonance for both bubble and solid particle suspensions. Existing experimental data for bubble distributions also shows a good agreement with the poly-disperse modifications proposed.

In this paper, the principles of Thickness-Expansion Mode (TEM) resonators for the characterization of fluids are described. From the measurement of the resonance parameters of a TEM piezoelectric transducer, the compressional acoustic impedance of gases and liquids can be determined. Since the propagation of mechanical waves into the fluid is not necessary, information in a wide range of frequencies can be obtained. Alternatively, these sensors can be driven in combination with other ultrasonic techniques to simultaneously determine the density, speed of sound and viscosity of samples. Some potential applications include the probe monitoring of processes and the characterization of fluids under harsh conditions. The main experimental criteria for the design and construction of high-resolution impedance meters (such as piezoelectric material, protective coating or thermal response) have been studied using equivalent electrical circuit modeling and finite element analysis., (Copyright © 2012 Elsevier B.V. All rights reserved.)

In this work, a high-resolution imaging method for the inspection of isotropic plate-like structures using linear arrays and Lamb waves is proposed. The evaluation of these components is limited by the low dynamic range resulting from main lobe and side lobe field patterns, and from the narrowband nature of the Lamb waves. Based on a full matrix array, synthetic aperture technique using all emitter-receiver combinations, different images from the same object are obtained by using different apodization coefficients, which are related to a trade-off between main lobe width and relative side lobe level. Several image compounding strategies have been tested and a new algorithm, based on apodization and polarity diversities between signals, is proposed. However, some effects, such as the dead zone close to the array and reverberations caused by interactions of the wavefront and defects, still limit the quality of the images. The use of spatial diversity, obtained by an additional array, introduces complementary information about the defects and improves the results of the proposed algorithm, producing high-resolution, high-contrast images. Experimental results are shown for a 1-mm-thick isotropic aluminum plate with artificial defects using linear arrays formed by 30 piezoelectric elements, with the low dispersion symmetric mode S0 at the frequency of 330 kHz.

A low-intensity ultrasonic technique (that is noninvasive, nondestructive, and online) has been developed to monitor the growth of Escherichia coli in glucose minimal media under aerobic and anaerobic conditions. Ultrasonic time of flight (TOF) variations were correlated with microorganism growth and the disappearance of nutrients and their subsequent conversion into different metabolites. Spectrophotometric growth data and high-performance liquid chromatography (HPLC) analysis of released and consumed metabolites were compared with the ultrasonic data demonstrating that the ultrasound device presented can provide supplementary real-time noninvasive information about the metabolic processes taking place in the culture. A semiempirical model under aerobic conditions was developed to explain the TOF variations as a function of the glucose concentration and was modified to take into account the growth inhibition due to the initial glucose concentration. The inhibition effect was obtained by fitting HPLC measurements to the logistic function of Boltzmann. Under aerobic and anaerobic culture conditions, the metabolic processes were correlated with TOF experimental variations through a theoretical approach based on ultrasonic propagation in ternary mixtures.

Milk contaminated with melamine resulted in an important health hazard that affected many babies in China recently. Ultrasonic characterization of adulterated milk may detect gross levels of melamine contamination. Sound speed and density measurements were made in skim milk as a function of melamine adulteration. An ultrasonic measurement technique to implement milk quality control is discussed.

In this work, fundamental aspects on the ultrasonic velocity monitoring of alcoholic fermentations in synthetic broths (glucose, fructose and sucrose) and natural media (must and wort) are reported. Results are explained in terms of monosaccharide catabolism, polysaccharide hydrolysis, gas production and microorganism growth. The effect of each one of these subprocesses upon ultrasonic velocity has been independently studied. It is shown that, regarding the sound propagation, the simplest systems behave as ternary dissolutions of sugar and ethanol in water, where, in the course of time, substrates are transformed into metabolites according to the fermentation reaction. A semi-empirical approach, based on the excess volume concept and the density and velocity measurements of binary mixtures, has been used to calculate these magnitudes in the ternary mixtures and to obtain the concentrations of the main solutes throughout the fermentations, reaching a good correlation (especially for the media of simplest composition). In all the processes analyzed, the data obtained from the ultrasonic measurements followed the changes caused by the yeast metabolism, asserting the potential of mechanical waves to monitor fermentations and, in general, biotechnological processes.

The use of a low-intensity ultrasonic technique (noninvasive, nondestructive, on-line, and able to assess opaque samples) to monitor the kinetics of invertase hydrolysis is presented. Adiabatic compressibility has been shown to be sensitive to sugar species: ultrasonic velocity increasing as saccharose is transformed into glucose and fructose. The influence of initial sucrose mass concentration (2-60%), temperature (25-55 degrees C), pH (3.5-6.5), and number of microorganisms (10(5)-10(9) yeasts/ml) on the reaction rate, catalyzed by the extracellular invertases of intact Saccharomyces cerevisiae cells, has been measured. The results were proven to be in strict agreement with the optimal kinetic parameters of the enzyme. Ultrasonic velocity variations are explained in terms of changes of the solute concentrations in the mixture water-saccharose-glucose/fructose and calculated from the velocity of ultrasound in the corresponding pure sugar solutions. A linear relationship between the initial rate of ultrasonic velocity and the number of yeasts (enzymes) is pointed out.