Ultrasonic measurement of temperature distributions in extreme environments: Electrical power plants testing in utility-scale steam generators.

Thermal heterogeneities within energy conversion and storage, material processing, nuclear processes, aerospace, and military applications are often inaccessible to characterization by insertion sensors. When sensor deployment is possible, conventional pointwise temperature probes quickly degrade when inserted into harsh environments typical of such processes. We developed spatially-resolved ultrasonic thermometry to noninvasively measure the spatial distributions of thermal properties in such applications, even when sizable thermal gradients are present. Our method divides the path of ultrasonic propagation into segments bound by echogenic features, which create echoes in pulse-echo mode, encoding the information about interior temperature distributions. We use the acquired ultrasonic responses to estimate the internal temperature distributions by solving an inverse problem or concatenating segmental estimates. This work describes the implementation and industrial testing of the developed method at a coal-fired electrical power generation plant. We inserted an echogenically segmented Inconel 625 waveguide into the combustion zone of the utility-scale boiler and continuously acquired ultrasonic data while keeping sensitive components away from the damaging combustion environment. The accuracy of the time-dependent temperature distributions reconstructed from the ultrasonic measurements was comparable to that of thermocouples. The resiliency of ultrasonic thermometry to harsh combustion conditions was far superior to conventional insertion sensors. The measurements obtained during plant operation captured daily steam generation cycles in response to changing customer demand and intermittent contributions of renewable power sources to the power grid. These measurements have revealed new insights into the relationship between the dynamic power generation load and the conditions inside the steam generator. The successful industrial testing of spatially-resolved ultrasonic thermometry in solids indicates that the developed technology has matured to become an attractive alternative to conventional sensing in solving challenging problems of long-term thermal characterizations in extreme environments.

Asymmetric depth-filtration: A versatile and scalable method for high-yield isolation of extracellular vesicles with low contamination.

We developed a novel asymmetric depth filtration (DF) approach to isolate extracellular vesicles (EVs) from biological fluids that outperforms ultracentrifugation and size-exclusion chromatography in purity and yield of isolated EVs. By these metrics, a single-step DF matches or exceeds the performance of multistep protocols with dedicated purification procedures in the isolation of plasma EVs. We demonstrate the selective transit and capture of biological nanoparticles in asymmetric pores by size and elasticity, low surface binding to the filtration medium, and the ability to cleanse EVs held by the filter before their recovery with the reversed flow all contribute to the achieved purity and yield of preparations. We further demonstrate the method's versatility by applying it to isolate EVs from different biofluids (plasma, urine, and cell culture growth medium). The DF workflow is simple, fast, and inexpensive. Only standard laboratory equipment is required for its implementation, making DF suitable for low-resource and point-of-use locations. The method may be used for EV isolation from small biological samples in diagnostic and treatment guidance applications. It can also be scaled up to harvest therapeutic EVs from large volumes of cell culture medium.

Quantification of Desiccated Extracellular Vesicles by Quartz Crystal Microbalance.

Extracellular vesicle (EV) quantification is a procedure through which the biomedical potential of EVs can be used and their biological function can be understood. The number of EVs isolated from cell culture media depends on the cell status and is especially important in studies on cell-to-cell signaling, disease modeling, drug development, etc. Currently, the methods that can be used to quantify isolated EVs are sparse, and each have limitations. In this report, we introduce the application of a quartz crystal microbalance (QCM) as a biosensor for quantifying EVs in a small drop of volatile solvent after it evaporates and leaves desiccated EVs on the surface of the quartz crystal. The shifts in the crystal's resonant frequency were found to obey Sauerbrey's relation for EV quantities up to 6 × 107, and it was determined that the biosensors could resolve samples that differ by at least 2.7 × 105 EVs. A ring-shaped pattern enriched in EVs after the samples had dried on the quartz crystal is also reported and discussed. QCM technology is highly sensitive and only requires small sample volumes and is significantly less costly compared with the approaches that are currently used for EV quantification.

Dynamic surface tension probe for measuring the concentration of extracellular vesicles.

The concentration of extracellular vesicles (EVs) is an essential attribute of biofluids and EV preparations. EV concentration in body fluids was correlated with health status. The abundance of EV secreted by cultured cells into growth medium is vital in signaling studies, tissue and disease models, and biomanufacturing of acellular therapeutic secretome. A limited number of physical principles sensitive to EV concertation have been discovered so far. Particle-by-particle counting methods enumerate individual particles scattering light, modulating the Coulter current, or appearing in EM images. The available ensemble techniques in current use rely on the concentration-dependent signal intensity, as in the case of ELISA. In this study, we propose for the first-time the ensemble-based characterization of EV concentration by dynamic surface tension (DST) probe and demonstrate its implementation. We show that DST measurements agree with the widely used NTA measurements of EV concertation. The proposed method is low-cost and requires only basic laboratory equipment for implementation.

Imaging of Extracellular Vesicles by Atomic Force Microscopy.

Exosomes and other extracellular vesicles (EVs) are molecular complexes consisting of a lipid membrane vesicle, its surface decoration by membrane proteins and other molecules, and diverse luminal content inherited from a parent cell, which includes RNAs, proteins, and DNAs. The characterization of the hydrodynamic sizes of EVs, which depends on the size of the vesicle and its coronal layer formed by surface decorations, has become routine. For exosomes, the smallest of EVs, the relative difference between the hydrodynamic and vesicles sizes is significant. The characterization of vesicles sizes by the cryogenic transmission electron microscopy (cryo-TEM) imaging, a gold standard technique, remains a challenge due to the cost of the instrument, the expertise required to perform the sample preparation, imaging and data analysis, and a small number of particles often observed in images. A widely available and accessible alternative is the atomic force microscopy (AFM), which can produce versatile data on three-dimensional geometry, size, and other biophysical properties of extracellular vesicles. The developed protocol guides the users in utilizing this analytical tool and outlines the workflow for the analysis of EVs by the AFM, which includes the sample preparation for imaging EVs in hydrated or desiccated form, the electrostatic immobilization of vesicles on a substrate, data acquisition, its analysis, and interpretation. The representative results demonstrate that the fixation of EVs on the modified mica surface is predictable, customizable, and allows the user to obtain sizing results for a large number of vesicles. The vesicle sizing based on the AFM data was found to be consistent with the cryo-TEM imaging.

Membrane proteins significantly restrict exosome mobility.

Exosomes are membrane nanovesicles implicated in cell-to-cell signaling in which they transfer their molecular cargo from the parent to the recipient cells. This role essentially depends on the exosomes' small size, which is the prerequisite for their rapid migration through the crowded extracellular matrix and into and out of circulation. Here we report much lower exosome mobility than expected from the size of their vesicles, implicate membrane proteins in a substantially impeded rate of migration, and suggest an approach to quantifying the impact. The broadly distributed excess hydrodynamic resistance provided by surface proteins produces a highly heterogeneous and microenvironment-dependent hindrance to exosome mobility. The implications of the findings on exosome-mediated signaling are discussed.

Ultrasound measurements of segmental temperature distribution in solids: Method and its high-temperature validation.

A novel approach that uses noninvasive ultrasound to measure the temperature distribution in solid materials is described and validated in high-temperature laboratory experiments. The approach utilizes an ultrasound propagation path with naturally occurring or purposefully introduced echogenic features that partially redirect the energy of an ultrasound excitation pulse back to the transducer, resulting in a train of echoes. Their time of flight depends on the velocity of ultrasound propagation, which changes with temperature distribution in different segments of the propagation path. We reconstruct segmental temperature distributions under different parameterizations. Several parameterizations are discussed, including piecewise constant and piecewise linear, and the parametrization that requires that the estimated temperature profile satisfies an appropriate heat conduction model. The experimental validation of the proposed approach with an alumina sample shows that even with simple parameterizations, the temperature profile is correctly captured with an accuracy that may be comparable to that of the traditional pointwise sensors. The advantages of the approach are discussed, including its suitability for real time and non-destructive temperature measurements in extreme environments and locations inaccessible to the traditional insertion sensors.

Size and shape characterization of hydrated and desiccated exosomes.

Exosomes are stable nanovesicles secreted by cells into the circulation. Their reported sizes differ substantially, which likely reflects the difference in the isolation techniques used, the cells that secreted them, and the methods used in their characterization. We analyzed the influence of the last factor on the measured sizes and shapes of hydrated and desiccated exosomes isolated from the serum of a pancreatic cancer patient and a healthy control. We found that hydrated exosomes are close-to-spherical nanoparticles with a hydrodynamic radius that is substantially larger than the geometric size. For desiccated exosomes, we found that the desiccated shape and sizing are influenced by the manner in which drying occurred. Isotropic desiccation in aerosol preserves the near-spherical shape of the exosomes, whereas drying on a surface likely distorts their shapes and influences the sizing results obtained by techniques that require surface fixation prior to analysis.

Surface tension of water in the presence of perfluorocarbon vapors.

Fluorocarbons are highly hydrophobic, biocompatible compounds with a variety of medical applications. Despite significant interest, the study of interfacial properties of fluorocarbons in aqueous systems has received limited attention. In this study, we investigate the influence of perfluoropentane and perfluorohexane vapors on the surface tension of water at room temperature. The results show a substantial decrease in the surface tension of water in the presence of perfluorocarbon vapors. In the investigated range of partial pressures up to the saturation value, a linear correlation between the surface tension and the partial pressure was found. This suggests that an adsorbed perfluorocarbon layer is formed on the surface of water. For comparison, the effect of the perfluorocarbon vapor on the surface tension of methanol was also investigated and a similar dependence was observed. Our results indicate that the stability and dynamic transitions of fluorocarbon colloids, which may be dispersed under physiological conditions as microdroplets, bubbles, or their combination, are likely affected by the composition of liquid and gas phases.

Identification of reduced-order thermal therapy models using thermal MR images: theory and validation.

In this paper, we develop and validate a method to identify computationally efficient site- and patient-specific models of ultrasound thermal therapies from MR thermal images. The models of the specific absorption rate of the transduced energy and the temperature response of the therapy target are identified in the reduced basis of proper orthogonal decomposition of thermal images, acquired in response to a mild thermal test excitation. The method permits dynamic reidentification of the treatment models during the therapy by recursively utilizing newly acquired images. Such adaptation is particularly important during high-temperature therapies, which are known to substantially and rapidly change tissue properties and blood perfusion. The developed theory was validated for the case of focused ultrasound heating of a tissue phantom. The experimental and computational results indicate that the developed approach produces accurate low-dimensional treatment models despite temporal and spatial noises in MR images and slow image acquisition rate.

Identification of controlled-complexity thermal therapy models derived from magnetic resonance thermometry images.

Medical imaging provides information valuable in diagnosis, planning, and control of therapies. In this paper, we develop a method that uses a specific type of imaging--the magnetic resonance thermometry--to identify accurate and computationally efficient site and patient-specific computer models for thermal therapies, such as focused ultrasound surgery, hyperthermia, and thermally triggered targeted drug delivery. The developed method uses a sequence of acquired MR thermometry images to identify a treatment model describing the deposition and dissipation of thermal energy in tissues. The proper orthogonal decomposition of thermal images is first used to identify a set of empirical eigenfunctions, which captures spatial correlations in the thermal response of tissues. Using the reduced subset of eigenfunction as a functional basis, low-dimensional thermal response and the ultrasound specific absorption rate models are then identified. Once identified, the treatment models can be used to plan, optimize, and control the treatment. The developed approach is validated experimentally using the results of MR thermal imaging of a tissue phantom during focused ultrasound sonication. The validation demonstrates that our approach produces accurate low-dimensional treatment models and provides a convenient tool for balancing the accuracy of model predictions and the computational complexity of the treatment models.

Bidirectional power stroke by ncd kinesin.

Optical trapping experiments reveal details of molecular motor dynamics. In noisy data, temporal structure within the power stroke of motors can be analyzed by ensemble averaging, but this obscures infrequent subcategories of events. We have here developed an analysis method that uses Kalman filtering of measurements, model-based estimation of the power strokes produced by the motor head, and automatic event classification to discriminate between different types of motor events. This method was applied to optical trap measurements of power strokes of the Drosophila kinesin-14 ncd in a three-bead geometry. We found the majority of events to be consistent with the previously discovered minus-end directed power stroke of ncd, occurring with ATP binding. Unexpectedly, 30% of apparent power strokes were plus-directed and 6% of binding events did not terminate in a discernible stroke. Ensemble averaging for each event category revealed that plus- and minus-directed strokes have different size and occur at different instants within the ncd-MT attachment sequence.

Comparison of surfactants used to prepare aqueous perfluoropentane emulsions for pharmaceutical applications.

Perfluoropentane (PFP), a very hydrophobic, nontoxic, noncarcinogenic fluoroalkane, has generated much interest in biomedical applications, including occlusion therapy and controlled drug delivery. For most of these applications, the dispersion within aqueous media of a large quantity of PFP droplets of the proper size is critically important. Surprisingly, the interfacial tension of PFP against water in the presence of surfactants used to stabilize the emulsion has rarely, if ever, been measured. In this study, we report the interfacial tension of PFP in the presence of surfactants used in previous studies to produce emulsions for biomedical applications: polyethylene oxide-co-polylactic acid (PEO-PLA) and polyethylene oxide-co-poly-epsilon-caprolactone (PEO-PCL). Because both of these surfactants are uncharged diblock copolymers that rely on the mechanism of steric stabilization, we also investigate for comparison's sake the use of the small-molecule cationic surfactant cetyl trimethyl ammonium bromide (CTAB) and the much larger protein surfactant bovine serum albumin (BSA). The results presented here complement previous reports of the PFP droplet size distribution and will be useful for determining to what extent the interfacial tension value can be used to control the mean PFP droplet size.

Method for measuring thickness of dielectric films using microdielectric fringe-effect sensors.

A method for noninvasive thickness measurements of dielectric films using fringe-effect (FE) sensors is developed and experimentally validated. The fringing electrical field, created by electrodes microfabricated at the film substrate, depends on the film thickness and dielectric permittivity of the film under test (FUT). The unknown film thickness is estimated by matching the theoretical prediction of thickness-dependent sensor admittance with the measured value. In the case of FE sensors with spatially periodic, interdigitated electrode (IDE) configuration, the admittance prediction is simplified, which allows for the real-time measurements of changing thickness. The developed method can be used to continuously measure the changing dielectric permittivity of the FUT material, which makes it possible to determine the thickness of films of changing dielectric properties, caused by chemical or other transformations. The application of the developed method is demonstrated experimentally by measuring the thickness of silicon nitride film deposited in several increments on the quartz substrate of the IDE sensor. In the expected range of sensor sensitivity, the results show an excellent agreement with the independent thickness measurements.

Physiological control of blood pumps using intrinsic pump parameters: a computer simulation study.

Implantable flow and pressure sensors, used to control rotary blood pumps, are unreliable in the long term. It is, therefore, desirable to develop a physiological control system that depends only on readily available measurements of the intrinsic pump parameters, such as measurements of the pump current, voltage, and speed (in revolutions per minute). A previously proposed DeltaP control method of ventricular assist devices (VADs) requires the implantation of two pressure sensors to measure the pressure difference between the left ventricle and aorta. In this article, we propose a model-based method for estimating DeltaP, which eliminates the need for implantable pressure sensors. The developed estimator consists of the extended Kalman filter in conjunction with the Golay-Savitzky filter. The performance of the combined estimator-VAD controller system was evaluated in computer simulations for a broad range of physical activities and varying cardiac conditions. The results show that there was no appreciable performance degradation of the estimator-controller system compared to the case when DeltaP is measured directly. The proposed approach effectively utilizes a VAD as both a pump and a differential pressure sensor, thus eliminating the need for dedicated implantable pressure and flow sensors. The simulation results show that different pump designs may not be equally effective at playing a dual role of a flow actuator and DeltaP sensor.

Control of thermal therapies with moving power deposition field.

A thermal therapy feedback control approach to control thermal dose using a moving power deposition field is developed and evaluated using simulations. A normal tissue safety objective is incorporated in the controller design by imposing constraints on temperature elevations at selected normal tissue locations. The proposed control technique consists of two stages. The first stage uses a model-based sliding mode controller that dynamically generates an 'ideal' power deposition profile which is generally unrealizable with available heating modalities. Subsequently, in order to approximately realize this spatially distributed idealized power deposition, a constrained quadratic optimizer is implemented to compute intensities and dwell times for a set of pre-selected power deposition fields created by a scanned focused transducer. The dwell times for various power deposition profiles are dynamically generated online as opposed to the commonly employed a priori-decided heating strategies. Dynamic intensity and trajectory generation safeguards the treatment outcome against modelling uncertainties and unknown disturbances. The controller is designed to enforce simultaneous activation of multiple normal tissue temperature constraints by rapidly switching between various power deposition profiles. The hypothesis behind the controller design is that the simultaneous activation of multiple constraints substantially reduces treatment time without compromising normal tissue safety. The controller performance and robustness with respect to parameter uncertainties is evaluated using simulations. The results demonstrate that the proposed controller can successfully deliver the desired thermal dose to the target while maintaining the temperatures at the user-specified normal tissue locations at or below the maximum allowable values. Although demonstrated for the case of a scanned focused ultrasound transducer, the developed approach can be extended to other heating modalities with moving deposition fields, such as external and interstitial ultrasound phased arrays, multiple radiofrequency needle applicators and microwave antennae.

MR thermometry-based feedback control of efficacy and safety in minimum-time thermal therapies: phantom and in-vivo evaluations.

The experimental validation of a model-based, thermal therapy control system which automatically and simultaneously achieves the specified efficacy and safety objectives of the treatment is reported. MR-thermometry measurements are used in real-time to control the power of a stationary, focused ultrasound transducer in order to achieve the desired treatment outcome in minimum time without violating the imposed safety constraints. Treatment efficacy is quantified in terms of the thermal dose delivered to the target. Normal tissue safety is ensured by automatically maintaining normal tissue temperature below the imposed limit in the user-specified locations. To reflect hardware limitations, constraints on the maximum applied power are also imposed. At the pretreatment stage, MR imaging and thermometry are used to localize the treatment target and identify thermal and actuation models. The results of phantom and canine experiments demonstrate that spatially-distributed, real-time MR temperature measurements enhance one's ability to robustly achieve the desired treatment outcome in minimum time without violating safety constraints. Post-treatment evaluation of the outcome using T2-weighted images of canine muscle showed good spatial correlation between the sonicated area and thermally damaged tissue.

Direct thermal dose control of constrained focused ultrasound treatments: phantom and in vivo evaluation.

The first treatment control system that explicitly and automatically balances the efficacy and safety goals of noninvasive thermal therapies is described, and its performance is evaluated in phantoms and in vivo using ultrasound heating with a fixed, focused transducer. The treatment efficacy is quantified in terms of thermal dose delivered to the target. The developed feedback thermal dose controller has a cascade structure with the main nonlinear dose controller continuously generating the reference temperature trajectory for the secondary, constrained, model predictive temperature controller. The control system ensures thermal safety of the normal tissue by automatically complying with user-specified constraints on the maximum allowable normal tissue temperatures. To reflect hardware limitations and to prevent cavitation, constraints on the maximum transducer power can also be imposed. It is shown that the developed controller can be used to achieve the minimum-time delivery of the desired thermal dose to the target without violating safety constraints, which is a novel and clinically desirable feature. The developed controller is model based, and requires patient- and site-specific models for its operation. These models were obtained during pre-treatment identification experiments. In our implementation, predictive models, internally used by the automatic treatment controller, are dynamically updated each time new temperature measurements become available. The adaptability of internal models safeguards against adverse effects of modelling errors, and ensures robust performance of the control system in the presence of a priori unknown treatment disturbances. The successful validation with two experimental models of considerably different thermal and ultrasound properties suggests the applicability of the developed treatment control system to different anatomical sites.

Minimum-time thermal dose control of thermal therapies.

The problem of controlling noninvasive thermal therapies is formulated as the problem of directly controlling thermal dose of the target. To limit the damage to the surrounding normal tissue, the constraints on the peak allowable temperatures in the selected spacial locations are imposed. The developed controller has a cascade structure with a linear, constrained, model predictive temperature controller in the secondary loop. The temperature controller manipulates the intensity of the ultrasound transducer with saturation constraints, which noninvasively heats the spatially distributed target. The main nonlinear thermal dose controller dynamically generates the reference temperature trajectories for the temperature controller. The thermal dose controller is designed to force the treatment progression at either the actuation or temperature constraints, which is required to minimize the treatment time. The developed controller is applicable to high and low-intensity treatments, such as thermal ablation and thermoradiotherapy. The developed approach is tested using computer simulations for a one-dimensional model of a tumor with constraints on the maximum allowable temperature in the normal tissue and a constrained power output of the ultrasound transducer. The simulation results demonstrate that the proposed approach is effective at delivering the desired thermal dose in a near minimum time without violating constraints on the maximum allowable temperature in healthy tissue, despite significant plant-model mismatch introduced during numerical simulation. The results of in vitro and in vivo validation are reported elsewhere.

Standard-independent estimation of dielectric permittivity with microdielectric fringe-effect sensors.

Microdielectric spectroscopy with planar fringe-effect (FE) interdigital sensors is a useful method for noninvasive characterization of the interfacial properties of the materials. Unfortunately, obtaining an accurate dielectric spectrum is difficult because of the complexity of the probing electrical field created by the FE sensor and the contribution of the sensor substrate and stray elements to the overall measurements. Previously, quantitative microdielectric spectroscopy required the calibration of the FE sensor with standard materials that are known to be dielectrically similar to an unknown sample of interest. This limitation complicates the application of microdielectric spectroscopy, particularly in cases where the monitored sample undergoes a transformation that changes its dielectric permittivity. A standard-independent method for quantitative FE microdielectric measurements is proposed in this paper. The developed method is based on comparison of the theoretically predicted admittance of the FE sensor with the sample of known dielectric properties and the measured sensor admittance. Comparison of the theoretical predictions with the admittance measurements reveals the contribution of the unknown stray elements. The measurements with an unknown sample are then adjusted for the strays. The contribution of the sensor substrate to the sensor measurements is removed using the theoretical model derived from the electroquasistatic approximation of Maxwell equations. The dielectric permittivity of the material being tested is calculated by successively solving the system of complex nonlinear equations for each frequency at which the sensor admittance is measured. The developed method is illustrated by applying it to the dielectric measurements of several dissimilar samples. The results are in excellent agreement with those obtained using the gold standard parallel-plate measurement method over the entire range of frequencies.