Reliable Method for Fabricating Tissue‐Mimicking Materials With Designated Relative Permittivity and Conductivity at 128 MHz

Artificial materials that can simultaneously mimic the relative permittivity and conductivity of various human tissues are usually used in medical applications. However, the method of precisely designing these materials with designated values of both relative permittivity and conductivity at 3 T MRI resonance frequency is lacking. In this study, a reliable method is established to determine the compositions of artificial dielectric materials with designated relative permittivity and conductivity at 128 MHz. Sixty dielectric materials were produced using oil, sodium chloride, gelatin, and deionized water as the main raw materials. The dielectric properties of these dielectric materials were measured using the open‐ended coaxial line method at 128 MHz. Nonlinear least‐squares Marquardt–Levenberg algorithm was used to obtain the formula, establishing the relationship between the compositions of the dielectric materials and their dielectric properties at 128 MHz. The dielectric properties of the blood, gall bladder, muscle, skin, lung, and bone at 128 MHz were selected to verify the reliability of the obtained formula. For the obtained formula, the coefficient of determination and the expanded uncertainties with a coverage factor of k = 2 were 0.991% and 4.9% for relative permittivity and 0.992% and 6.4% for conductivity. For the obtained artificial materials measured using the open‐ended coaxial line method, the maximal difference of relative permittivity and conductivity were 1.0 and 0.02 S/m, respectively, with respect to the designated values. In conclusion, the compositions of tissue‐mimicking material can be quickly determined after the establishment of the formulas with the expanded uncertainties of less than 10%. Bioelectromagnetics. 2021;42:86–94. © 2020 Bioelectromagnetics Society.     

Quantitative modeling of viable cell density,cell size,intracellular conductivity,and membrane capacitance in batch and fed‐batch CHO processes using dielectric spectroscopy

Dielectric spectroscopy was used to analyze typical batch and fed‐batch CHO cell culture processes. Three methods of analysis (linear modeling, Cole–Cole modeling, and partial least squares regression), were used to correlate the spectroscopic data with routine biomass measurements [viable packed cell volume, viable cell concentration (VCC), cell size, and oxygen uptake rate (OUR)]. All three models predicted offline biomass measurements accurately during the growth phase of the cultures. However, during the stationary and decline phases of the cultures, the models decreased in accuracy to varying degrees. Offline cell radius measurements were unsuccessfully used to correct for the deviations from the linear model, indicating that physiological changes affecting permittivity were occurring. The β‐dispersion was analyzed using the Cole–Cole distribution parameters Δε (magnitude of the permittivity drop), f_c (critical frequency), and α (Cole–Cole parameter). Furthermore, the dielectric parameters static internal conductivity (σ_i) and membrane capacitance per area (C_m) were calculated for the cultures. Finally, the relationship between permittivity, OUR, and VCC was examined, demonstrating how the definition of viability is critical when analyzing biomass online. The results indicate that the common assumptions of constant size and dielectric properties used in dielectric analysis are not always valid during later phases of cell culture processes. The findings also demonstrate that dielectric spectroscopy, while not a substitute for VCC, is a complementary measurement of viable biomass, providing useful auxiliary information about the physiological state of a culture. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010     

Complex Dielectric Properties of Sulfate-Reducing Bacteria Suspensions

Sulfate-reducing bacteria (SRB) can potentially enhance the remediation of heavy metals in the subsurface. Previous geophysical research has demonstrated the sensitivity of electrical measurements to SRB-mediated mineral transformation in porous media. However, the inherent dielectric properties of SRB and their direct contribution to the electrical properties of porous media are poorly understood. We studied the complex dielectric properties of SRB (Desulfovibrio vulgaris) suspensions at different concentrations and at different growth stages using a two-electrode dielectric spectroscopy measurement over the frequency range of 20 Hz to 1 MHz. Our results show higher dielectric responses (relative dielectric permittivity, real and imaginary conductivity) occurred with higher bacteria concentration at frequencies <10 kHz. Additionally, permittivity and conductivity both decreased as cells aged from mid-log phase to late stationary phase. Our results suggest that dielectric spectroscopy measurements can be used to noninvasively monitor biomass and various growth stages of SRB. Our work advances the interpretation of electrical signals associated with SRB observed in the subsurface.     

A novel approach for using dielectric spectroscopy to predict viable cell volume (VCV) in early process development

Online monitoring of viable cell volume (VCV) is essential to the development, monitoring, and control of bioprocesses. The commercial availability of steam‐sterilizable dielectric‐spectroscopy probes has enabled successful adoption of this technology as a key noninvasive method to measure VCV for cell‐culture processes. Technological challenges still exist, however. For some cell lines, the technique's accuracy in predicting the VCV from probe‐permittivity measurements declines as the viability of the cell culture decreases. To investigate the cause of this decrease in accuracy, divergences in predicted vs. actual VCV measurements were directly related to the shape of dielectric frequency scans collected during a cell culture. The changes in the shape of the beta dispersion, which are associated with changes in cell state, are quantified by applying a novel “area ratio” (AR) metric to frequency‐scanning data from the dielectric‐spectroscopy probes. The AR metric is then used to relate the shape of the beta dispersion to single‐frequency permittivity measurements to accurately predict the offline VCV throughout an entire fed‐batch run, regardless of cell state. This work demonstrates the possible feasibility of quantifying the shape of the beta dispersion, determined from frequency‐scanning data, for enhanced measurement of VCV in mammalian cell cultures by applying a novel shape‐characterization technique. In addition, this work demonstrates the utility of using changes in the shape of the beta dispersion to quantify cell health. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 30:479–487, 2014     

Dielectric Properties Based Detection of Heavy Metal Contaminated Soil in the Frequency Range from 10 MHz to 1 GHz

Physical parameters based electromagnetic methods are promising technologies to detect contaminated sites. In these methods, the dielectric property is a key parameter. In this paper, we studied the dielectric characteristics of heavy metal contaminated soil. The chromium contaminated soil was made into samples, and the open-ended coaxial line was adopted as the measurement method. Experiments were conducted in the frequency band between 10 MHz and 1 GHz. The results showed that the complex permittivity, including the real part and the imaginary part, changes as the ionic content changes. Especially, at low frequencies (<50 MHz), the complex permittivity increases significantly with the increase of ionic content. In addition, it also could be seen that the water content of the soil also affects the complex permittivity. We proposed to adopt the drying method or the Time Domain Reflection method to determine the water content. The dielectric parameters are most affected by the ionic content after knowing the water content. Therefore, it is feasible to detect heavy metal contaminated sites based on dielectric properties.     

生物量浓度实时在线检测方法的研究

微生物的存在会改变发酵液的电特性,发酵液在无线电频率范围内的电容率增量是测量频率和生物量浓度的函数.基于对发酵液电容率分布的研究,提出了测量生物量浓度的新方法.用此方法不用取样就能对发酵液中的生物量进行实时在线测量,而且测得的是活的生物量浓度.制作的电极直接插入发酵器中并满足高温蒸气灭菌条件.此方法在生化制药、食品发酵、啤酒酿造、污水检测等工业领域里有很好的推广应用前景.     

Dielectric and conformal studies of 1-propanol and 1-butanol in methanol

Hydrogen bonds in small chain alcohol-alcohol binary systems alter the dielectric permittivity of the binary system. With a view to obtain a better understanding of the interactions in such systems, the complex permittivity spectra of mixtures of methanol (ME) with 1propanol (1PR) and 1butanol (1BU) have been measured in the frequency range 10 MHz to 20 GHz using time domain reflectometry at 288 K, 298 K, 308 K and 318 K. The dielectric parameters such as static dielectric constant and relaxation time were obtained using the calibration method based on nonlinear least squares fit method. Using these parameters excess permittivity, excess inverse relaxation time, Kirkwood correlation factor, and thermodynamic parameters were determined. It is observed that the static permittivity decreases with increase in mole fractions of 1PR/1BU in ME whereas the relaxation time increases for both the binary systems. Computational conformational analysis was performed using ab initio Hartree-Fock using Gaussian-03 program. The present studies indicate a difference in the solvation of ME by 1PR/1BU and vice versa. Further, the interaction of ME-1PR is distinctly different at the 0.55 molar concentration of 1PR while the ME-1BU system shows strong interactions in both the methanol and the 1BU rich regions.     

Ab initio and dielectric studies of succinic acid and maleic acid in 1,4-dioxane

In a continuing effort to understand the hydrogen bond through the study of dielectric and computational conformal studies of dilute solutions, succinic acid and maleic acid are studied in solutions of 1,4-dioxane solvent. Dielectric studies give an account of the net dipole moment of the system under study, which is then compared with the values obtained from conformal analysis. The dielectric measurements were made at 303 K at a frequency of 9.83 GHz using a X-band microwave test bench in order to determine the relaxation times and the dipole moments. The static dielectric permittivity and the high frequency dielectric permittivity were measured using a LCR meter and an Abbe's refractometer, respectively. The results are inspected in comparison with the dipole moment results of ab initio calculations of some of the conformers in gas phase and in liquid phase. Gaussian-03 software package with 6–31G(d) basis set optimisation was used for this purpose. Onsager's reaction field model is used to study the solvation of the dicarboxylic acids in 1,4-dioxane. The results are interpreted in terms of the intermolecular and intramolecular hydrogen bond interactions in the dilute systems.     

Evaluation of leaf water status by means of permittivity at terahertz frequencies

We present an electromagnetic model of plant leaves which describes their permittivity at terahertz frequencies. The complex permittivity is investigated as a function of the water content of the leaf. Our measurements on coffee leaves (Coffea arabica L.) demonstrate that the dielectric material parameters can be employed to determine the leaf water status and, therefore, to monitor drought stress in plant leaves. The electromagnetic model consists of an effective medium theory, which is implemented by a third order extension of the Landau, Lifshitz, Looyenga model. The influence of scattering becomes important at higher frequencies and is modeled by a Rayleigh roughness factor.     

Analytical Approach to the Prism Coupling Problem in the Kretschmann Configuration

Prism coupling in the Kretschmann configuration is a well-known method for excitation of surface plasmon polaritons (SPP’s) in a metal film bounded from one side by a prism and from the other side by air. The analysis of the reflectance in the upper medium (prism) is based on the well-known Fresnel’s formula. Due to the fact that this formula cannot be inverted directly to give the complex dielectric permittivity and the thickness of the metal film from measured values of the reflectance at three or more different angles of incidence, an additional analysis is needed. Here, such an analysis is presented. The special case of illumination with He–Ne laser (λ = 632.8 nm) of silver film bounded by air is considered. A new asymptotic formula for the Lorentz dip is derived. Our experimental data for silver film are reported too.     

Real-time monitoring of yeast cell division by dielectric spectroscopy.

To assay cell cycle progression in synchronized culture of yeast we have applied dielectric spectroscopy to its real-time monitoring. The dielectric monitoring is based on the electromagnetic induction method, regarded as a nonelectrode method, which has resolved the problems encountered in measurements with metal electrodes, namely electrode polarization and bubble formation on electrodes. In the synchronized culture with temperature-sensitive cell division cycle mutants, the permittivity of the culture broth showed cyclic changes at frequencies below 300 kHz. The increase and decrease in the cyclic changes of the relative permittivity correspond to the increase in cell length and bud size and to the septum formation between mother and daughter cells, respectively.     

Substitution and spreadsheet methods for analysing dielectric spectra of biological systems

1. Two major problems are encountered when one wishes to fit audio- and radio-frequency dielectric spectra of biological cell suspensions (or other materials): (a) changes in the apparent frequency-dependent permittivity of the system due to the phenomena of electrode polarisation can dominate those due to the biological system, and (b) because of the overlap of different dispersions it may be very difficult to deconvolute the individual contributions of the underlying biophysical mechanisms. 2. The extent of electrode polarisation depends substantially upon the conductivity of the medium surrounding the cells, but only marginally on the nature of the ions of a given valency contribution to it. 3. This, and the fact that the apparent time constants of the phenomena contributing to electrode polarisation are much greater than those of biological dielectric dispersions, permits one to use a simple substitution method to extract the latter in the presence of the former. This is shown both by simulation and by experiments using suspensions of human erythrocytes. 4. A spreadsheet method is described for the display of dielectric data and their conformance to the double Cole-Cole equation. The method provides a rapid and convenient approach, based on interactive graphical outputs, for the fitting of dielectric data to this equation. 5. Estimates derived from the spreadsheet program may be used in a BASIC program to arrive at the optimal fit. 6. The method is applied to the strongly-overlapping - and -dispersions of erythrocytes, permitting their deconvolution and providing a high level of accuracy. Offprint requests to: D. B. Kell     

Dielectric monitoring of growth and sporulation of <Emphasis Type="BoldItalic">Bacillus thuringiensis</Emphasis>

On-line permittivity and optical density measurements have been used to monitor biomass concentration and sporulation status during growth of a spore-forming bacterium, Bacillus thuringiensis, in fed-batch culture. The correlation between permittivity, optical density and other observations showed three distinct phases of growth: growth itself, transition and sporulation. The permittivity variations during the transition and sporulation phases could be related to the sporulation development: the evolution pattern of the ratio of optical density to permittivity was representative of the culture state, and during the sporulation phase, a permittivity index could be build to measure the extend of spore liberation.     

Ions around DNA: Monte Carlo estimates of distribution with improved electrostatic potentials

Coulomb's law for the electrostatic interactions between ions is modified when discontinuities in dielectric constant (relative permittivity) occur. In a DNA solution such a discontinuity occurs at the interface between the DNA molecular helix and the surrounding water. We take the modified interaction potentials from a previous report [Macromolecules (1986) 19 , 1186–1194] and use them with the Monte Carlo method to find the distribution of univalent and bivalent counterions around the DNA helix in the absence of coions (i.e., no added salt). In comparing the ion distribution with the modified potential to the distribution without, we find that the effects of the modifications to the potentials are considerable. The modifications tend to drive the ions out of the grooves of the helix, especially out of the major groove. This result comes partly from the repulsion exerted on the ions by the low-permittivity helix and partly from the concentration of the field of the phosphates at the surface of the helix, a concentration that is also caused by the discontinuity in permittivity.     

Real-time monitoring of adherent Vero cell density and apoptosis in bioreactor processes

This study proposes an easy to use in situ device, based on multi-frequency permittivity measurements, to monitor the growth and death of attached Vero cells cultivated on microporous microcarriers, without any cell sampling. Vero cell densities were on-line quantified up to 10⁶ cell mL⁻¹. Some parameters which could potentially impact Vero cell morphological and physiological states were assessed through different culture operating conditions, such as media formulation or medium feed-harvest during cell growth phase. A new method of in situ cell death detection with dielectric spectroscopy was also successfully implemented. Thus, through permittivity frequency scanning, major rises of the apoptotic cell population in bioreactor cultures were detected by monitoring the characteristic frequency of the cell population, f_c, which is one of the culture dielectric parameters. Both cell density quantification and cell apoptosis detection are strategic information in cell-based production processes as they are involved in major events of the process, such as scale-up or choice of the viral infection conditions. This new application of dielectric spectroscopy to adherent cell culture processes makes it a very promising tool for risk-mitigation strategy in industrial processes. Therefore, our results contribute to the development of Process Analytical Technology in cell-based industrial processes.     

Dielectric measurement to monitor the growth and the physiological states of biological cells

Measurement of capacitance, also referred to as dielectric permittivity, is a new method of estimating the concentration of cells, monitoring the growth and detecting the physiological changes during the cultivation of organisms in various bioprocess. Several types of biological cells were studied, namely; Saccharomyces cerevisiae, Escherichia coli, Perilla frutescens (plant cells) and AFP-27 hybridoma cells. Generally, a linear correlation between cell capacitance (C) and other biomass measurement technique such as optical density (OD) and dry weight (DW) was obtained using the different types of cell suspension. Therefore, this method could be used to monitor the growth of the organism during the active growth. It could be conveniently used to make a rapid estimate of the cell concentration such as in plant cell suspension culture. The capacitance sensor could also be designed to be installed and autoclaved in-situ in a bioreactor and used for on-line monitoring of cell growth. On the other hand, distinct deviations in the capacitance value were observed in relation with the growth stage of the organism. This was observed in all the organisms studied but the type of deviation depends on the physiology of the organism. This variation in cell capacitance showed the possibility of using this method as a means to indicate changes in the physiological state of cells during cultivation. This capability would be very useful in designing control strategies that would depend on the physiological states in the bioprocess. Present address: Miles Inc., Berkeley, CA 94701 U.S.A.The authors sincerely appreciated the generosity of Dr. K. Mishima and Dr. A. Mimura of Kobe Steel Co., Japan. The useful discussions with M. Nakajima and technical assistance of J. Zhong and R. Pambayun were also acknowledged. The work in hybridoma cell culture was done through the collaboration with C. Perusich-Kussow and Prof. W. S. Hu, University of Minnesota, USA.     

Polymer Nanocomposites with Interpenetrating Gradient Structure Exhibiting Ultrahigh Discharge Efficiency and Energy Density

Poly(vinylidene fluoride) (PVDF) based polymer nanocomposites with high‐permittivity nanofillers exhibit outstanding dielectric energy storage performance due to their high dielectric permittivities and breakdown strength. However, their discharge efficiency is relatively low (usually lower than 70%), which limits their practical applications. Here, polymer nanocomposites with a novel interpenetrating gradient structure are designed and demonstrated by cofilling a PVDF matrix with barium zirconate titanate nanofibers and hexagonal boron nitride nanosheets via modified nonequilibrium processing. The interpenetrating gradient structure is highly effective in breaking the trade‐off between discharge energy density and efficiency of the corresponding nanocomposite, as indicated by the concomitantly enhanced discharge energy density (U _e ≈ 23.4 J cm^?3) and discharge efficiency (η ≈ 83%). The superior performance is primarily attributed to the rational distribution of nanofillers in the polymer matrix, which raises the height of the potential barrier for charge injection at the dielectric/electrode interface, suppresses electric conduction and contributes to enhanced apparent breakdown strength. Meanwhile, the gradient configuration allows higher volume fraction of high‐permittivity nanofillers without compromising the breakdown strength, leading to higher electric polarization compared with the random configuration. This work provides new opportunities to PVDF‐based polymer nanocomposites with high energy density and discharge efficiency for capacitive energy storage applications.     

Characterizing cellular systems by means of dielectric spectroscopy

The dielectric behavior of a suspension of synchronized, spherical cells has been investigated in relation to the electrical parameters of certain cell structures. In the quasistatic approximation, Poisson's equations are solved for the respective diffusive media, and the local charge distributions are derived by taking into account the continuity equations. The results describe both α and β dispersion and reduce, in the corresponding limiting cases, to previous reports. The dependence of suspension permittivity in α-and β-dispersion ranges on the diffusive effects, the conductivity, and the permittivity of cytoplasm, of membrane, and of culture medium as well as on membrane thickness is pointed out. The possibility is pointed out of characterizing cellular behavior by means of the evolution of certain electrical and morphological parameters during cell cycle progression as well the effects of different stimuli on cellular systems derived by fast dielectric spectroscopy. © 1996 Wiley-Liss, Inc.     

Microwave Resonant Sensor for Non-invasive Characterization of Biological Tissues

Context

Microwave sensing appears to be an open wide field to investigate medical applications, such as monitoring of vital signs (temperature, arterial pressure, …), following different kinds of pathologies (cancer, glucose level …) and aid for medical diagnosis. It offers an alternative to determine the dielectric properties of biological tissues through the development of local non-invasive and/or embedded sensors, giving thus a kind of imaging by the dielectric contrast. Moreover, RF communications links between several sensors can be developed to realize “Body Area Networks”.

Online- and offline- monitoring of stem cell expansion on microcarrier

In the biopharmaceutical industry, adherent growing stem cell cultures gain worldwide importance as cell products. The cultivation process of these cells, such as in stirred tank reactors or in fixed bed reactors, is highly sophisticated. Cultivations need to be monitored and controlled to guarantee product quality and to satisfy GMP requirements. With the process analytical technology (PAT) initiative, requirements regarding process monitoring and control have changed and real-time on-line monitoring tools are recommended. A tool meeting the new requirements may be the dielectric spectroscopy for online viable cell mass determination by measurement of the permittivity. To establish these tools, proper offline methods for data correlation are required. The cell number determination of adherent cells on microcarrier is difficult, as it requires cell detachment from the carrier, which highly increases the statistical error. As an offline method, a fluorescence assay based on SYBR^®GreenI was developed allowing fast and easy total cell concentration determination without the need to detach the cells from the carrier. The assay is suitable for glass carriers used in stirred tank reactor systems or in fixed bed systems, may be suitable for different cell lines and can be applied to high sample numbers easily. The linear dependency of permittivity to cell concentration of suspended stem cells with the dielectric spectroscopy is shown for even very small cell concentrations. With this offline-method, a correlation of the cell concentration grown on carrier to the permittivity data measured by the dielectric spectroscopy was done successfully.