[Show abstract][Hide abstract] ABSTRACT: Experimental evidence shows that nanosecond pulsed electric fields (nsPEF) trigger apoptosis in skin tumors. We have postulated that the energy delivered by nsPEF is insufficient to impart significant heating to the treated tissue. Here we use both direct measurements and theoretical modeling of the Joule heating in order to validate this assumption.
For the temperature measurement, thermo sensitive liquid crystals (TLC) were used to determine the surface temperature while a micro-thermocouple (made from 30 μm wires) was used for measuring the temperature inside the tissue. The calculation of the temperature distribution used an asymptotic approach with repeated calculation of the electric field, Joule heating and heat transfer, and subsequent readjustment of the electrical tissue conductivity. This yields a temperature distribution both in space and time.
It can be shown that for the measured increase in temperature an unexpectedly high electrical conductivity of the tissue would be required, which was indeed found by using voltage and current monitoring during the experiment. Using impedance measurements within tafter = 50 μs after the pulse revealed a fast decline of the high conductivity state when the electric field ceases. The experimentally measured high conductance of a skin fold (mouse) between plate electrodes was about 5 times higher than the maximally expected conductance due to fully electroporated membrane structures (Gmax/Gelectroporated) ≈ 5. Fully electroporated assumes that 100 % of the membranes are conductive which is estimated from impedance measurement at 10 MHz where membranes are capacitively shorted. Since the temperature rise in B-16 mouse melanoma tumors due to equally spaced (Δt = 2 s) 300 ns-pulses with E = 40 kV/cm usually does not exceed ΔΤ = 3 K at all parts of the skin fold between the electrodes, a hyperthermic effect on the tissue can be excluded.
[Show abstract][Hide abstract] ABSTRACT: Titanium (Ti) and its alloys are commonly used as implant materials. As a result of abrasion, micron- and nano-sized particles can be released into the tissues surrounding Ti-based implants. Previous studies have shown that nanoparticles (NPs) are more biologically active and cytotoxic than micron-sized particles. Objectives: The effect of titanium dioxide (TiO2) NPs on cellular proliferation, viability, morphology, spreading, and detachment from substrate was investigated. Methods: An impedance-based assay was chosen due to its advantages of being label-free and monitoring the cells in real-time. Trypan Blue exclusion and ultrahigh resolution imaging (URI) were also employed. Fibroblasts seeded in microelectrode-embedded E-plates were exposed to 0.05, 0.5, and 5 mg/L of nano-TiO2 for up to 120 h. The particles used (Sigma–Aldrich, Norway) had a diameter < 100 nm and a mixed anatase /rutile crystallographic form. An alternative excitation signal (20 mV control voltage amplitude) was applied at 10, 25, and 50 kHz to the microelectrodes in the E-plates. Cells act as insulators and their attachment to the electrode surfaces increases the impedance. Continuous impedance measurements were made with an xCELLigence system (ACEA Inc, USA). The experiments were run in duplicate and repeated at least three times. Unexposed cells served as controls. Results: Both the impedance-based monitoring and the Trypan blue assay showed an overall decrease of proliferation and viability of the cells exposed to TiO2 NPs. URI microscopy revealed a functional dependence of apoptosis on the nano-TiO2 concentration. Conclusion: Overall, impedance-based results were in accordance with those from the Trypan Blue exclusion and URI and indicated a significant cytotoxic effect at 5 mg/L of TiO2NPs.
The study was supported by funding from UH-nett Vest, the Department of Clinical Dentistry, Faculty of Medicine and Dentistry in Bergen, and the Regional Development Program Hordaland-Thüringen.
Annual Meeting of the IADR Continental European Division 2013; 09/2013
[Show abstract][Hide abstract] ABSTRACT: Numerous studies have shown electromyographic signals (EMGs) as useful for controlling prostheses and ortheses. Their great potential stems from the degree of voluntary control we wield over these signals, even if limbs are missing, especially in the EMGs of skeletal muscles. However, despite several decades of exploration, the potential of electrical myoimpedance (EMI) as a separate control signal has been largely ignored. The greatest barrier to utilizing this signal is that skin-based measurements suffer from low-sensitivity with the only alternative being invasive methods, such as needles.
Since the EMI signal is known to correlate with the passive properties of materials under test, specifically their geometry, it is expected to be highly sensitive to the morphologic changes which occur during concentric contractions. In contrast, EMG signals occur any time a muscle is contracted, whether or not this results in morphologic changes. Therefore, the EMI and EMG signals should not be highly-correlated, suggesting an additional control channel.
To test this, we developed a non-invasive procedure for making simultaneous skin-based measurements of EMG and EMI signals during both concentric and isometric contractions. A video camera was synchronized with the measurement system to facilitate the correlation of signal features with muscle actions. We conclude that the two signals can be distinguished.
15th International Conference on Electrical Bio-Impedance, Heilbad Heiligenstadt, Germany; 04/2013
[Show abstract][Hide abstract] ABSTRACT: Impedance-based assays can constitute a reliable alternative to the
conventional methods used in nanotoxicology due to the important
advantages of being label-free and monitoring the cells in real-time. In
this study, the suitability of impedance-monitoring for the screening of
nanoparticle (NP)-induced cytotoxicity was assessed. The effect of
titanium dioxide (TiO2)-NPs on cellular proliferation,
viability, spreading, and detachment from substrate was evaluated by
continuous impedance-based measurements made with an xCELLigence system.
Fibroblasts seeded in microelectrode-embedded E-plates were exposed to
spherical anatase nano-TiO2 (5, 10, and 40 nm in diameter)
for up to 120 h. An alternative excitation signal (20 mV control voltage
amplitude) was applied at 10, 25, and 50 kHz to the microelectrodes in
the E-plates. Cells attached to the electrode surfaces act as insulators
and lead to an increase in impedance. For validating the
impedance-method, Trypan Blue exclusion and ultrahigh resolution imaging
(URI) were employed. The general trend observed was a decrease in
impedance following exposure to TiO2-NPs. Impedance-based
results were in most instances in accordance with those from the Trypan
Blue exclusion and URI assays indicating that the impedance-based
approach has merit. Further studies are needed to validate it as a
high-throughput method for evaluating NPs' cytotoxicity.
Journal of Physics Conference Series 04/2013; 429(1). DOI:10.1088/1742-6596/429/1/012026
[Show abstract][Hide abstract] ABSTRACT: While most applications of bio-impedance measurements for
characterization of cells or tissue do not have any time requirements,
newer developments like real time monitoring of moving cells or membrane
recovery after high voltage application depend on fast measurement. This
is not compatible with sweeping the frequency through the desired range
while assessing magnitude and phase of the material under test (MUT). A
high speed is only achievable by using broad-bandwidth excitation
signals and monitoring the response in time-domain. Time-domain based
methods can be distinguished by the excitation signals as well as by
assessing the transmission or reflection behavior of the MUT.
Although there is good agreement regarding the advantages of fast
measurements, time-domain measurements are often rejected because of low
precision and noise sensitivity. This paper points not only the
advantages of impedance measurements in time domain but shows also
drawbacks together with possible solutions.
Journal of Physics Conference Series 04/2013; 434(1):2092-. DOI:10.1088/1742-6596/434/1/012092
[Show abstract][Hide abstract] ABSTRACT: This study presents the further establishment of impedimetric biosensors
with aptamers as receptors. Aptamers are short single-stranded
oligonucleotides which bind analytes with a specific region of their 3D
structure. Electrical impedance spectroscopy is a sensitive method for
analyzing changes on the electrode surface, e.g. caused by
receptor-ligand-interactions. Fast and inexpensive prototyping of
electrodes on the basis of commercially available compact discs having a
24 carat gold reflective layer was investigated. Electrode structures
(CDtrodes ) in the range from few millimetres down to 100 microns
were realized. The well-studied thrombin-binding aptamer (TBA) was used
as receptor for characterizing these micro- and macro-electrodes. The
impedance signal showed a linear correlation for concentrations of
thrombin between 1.0 nM to 100 nM. This range corresponds well with most
of the references and may be useful for the point-of-care testing
Journal of Physics Conference Series 04/2013; 434(1):2091-. DOI:10.1088/1742-6596/434/1/012091
[Show abstract][Hide abstract] ABSTRACT: Electrolytic conductivity of cell-based suspensions is an important
parameter which can be easily and non-destructively measured as part of
the electrical impedance. The low frequency conductivity of a cell
suspension with very low cell density equals nearly the medium
conductivity. However, a high cell density decreases the low frequency
conductivity due to the insulating behaviour of the cell membranes.
Here we use miniaturized electrode structures, smaller than the size of
typical cells for impedometric conductivity measurement which allows an
impedance measurement independent of the electrical properties of
suspended cells or particles.
Journal of Physics Conference Series 04/2013; 434(1):2093-. DOI:10.1088/1742-6596/434/1/012093
[Show abstract][Hide abstract] ABSTRACT: The AD5933 , a specialized single chip impedance analyzer, made by
Analog Devices, is basically not intended for use with four electrode
interface. Due to electrochemical phenomena at the electrodes connecting
the material under test (MUT), especially in the low frequency region
below 100 kHz, a two electrode interface generates considerable errors
during the measurement. Thus, for most application in bio-impedance
measurement only a four electrode interface can guarantee reliable
results. Here we show how a four electrode interface with galvanostatic
excitation but also for potentiostatic excitation can be realized by
just a few external components.
Journal of Physics Conference Series 12/2012; 407(1):2019-. DOI:10.1088/1742-6596/407/1/012019
[Show abstract][Hide abstract] ABSTRACT: Electrodes are an important part of the impedance measurement chain but
their influence is often underestimated. Electrochemical reactions or
polarization effects, especially for galvanically coupled systems are
sometimes completely neglected. The most important features of
electrodes to be considered are the geometry with respect to the
structure of the material to be tested and the electrochemistry at the
electrode surface. Especially drift and corrosion effects may yield
misleading results. If applicable, sophisticated electrode systems
should be used in order to prevent the distorting influence of electrode
polarization, for extending the useful frequency range of the electrodes
or for enhancing the signal-to-noise ratio.
Journal of Physics Conference Series 12/2012; 407(1):2027-. DOI:10.1088/1742-6596/407/1/012027
[Show abstract][Hide abstract] ABSTRACT: The incorporation of photoactive molecules in thin layers enables photoinduced changes in wettability, e.g. azobenzenes, or micropatterning by deprotection of functional groups, e.g. applying nitroveratryl compounds. This paper describes a synthetic route for obtaining an azobenzene with a silane anchor. The chemical synthesis, including all intermediates, is characterized by NMR and IR. The photoisomerization of all products was investigated by UV/Vis spectroscopy. Ellipsometry and contact angle measurements give information about monolayers of the synthesized organosilanes.
[Show abstract][Hide abstract] ABSTRACT: Proper illumination is essential for light microscopy. Whereas in early years incandescent light was the only illumination, today, more and more specialized light sources, such as lasers or arc lamps are used. Because of the high efficiency and brightness that light-emitting diodes (LED) have reached today, they have become a serious alternative for almost all kinds of illumination in light microscopy. LED have a high durability, do not need expensive electronics, and they can be switched in nanoseconds. Besides this, they are available throughout the UV/Vis/NIR-spectrum with a narrow bandwidth. This makes them ideal light sources for fluorescence microscopy. The white LED, with a color temperature ranging from 2,600 up to 5,000 K is an excellent choice for bright-field illumination with the additional advantage of simple brightness adjustments without changing the spectrum. This review discusses the different LED types, their use in the fluorescence microscope, and discusses LED as specialized illumination sources for Förster resonance energy transfer and fluorescent lifetime imaging microscopy.
Cytometry Part A 03/2012; 81(3):188-97. DOI:10.1002/cyto.a.22023 · 2.93 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Square wave excitation is an excellent choice for excitation signal in broad bandwidth and fast impedance measurement. The advantages are simple scalability, easy generation and straight forward data reduction by non-uniform sampling. The often cited disadvantage of low signal energy at high frequency is only partially relevant because a high sampling rate at fast changing portions of the measured signal guarantees a sufficient signal-to-noise ratio throughout the frequency range determined by the sampling speed, the slew rate of the amplifiers used and the time duration of the excitation signal. Although briefly looking at spectra of other signals suggests their better performance, having a well-designed system from signal generation through sampling and processing up to proper data mining shows the advantages of square wave excitation, especially if speed and bandwidth is important. Here we describe a method for obtaining an estimate of a broadband spectrum over several orders of magnitude while only requiring a minimum of computing power.
[Show abstract][Hide abstract] ABSTRACT: Fast impedance measurements are often performed in time domain utilizing broad bandwidth excitation signals. Other than in frequency domain measurements harmonic distortion cannot be compensated which requires careful design of the analog front end. In order to minimize the influence of electrode polarization and noise, especially in low-frequency measurements, current injection shows several advantages compared to voltage application. Here, we show an active front end based on a voltage-controlled current source for a wide range of impedances. Using proper feedback, the majority of the parasitic capacitances are compensated. The bandwidth ranges from dc to 20 MHz for impedance magnitude below 5 kΩ. The output is a symmetric signal without dc-offset which is accomplished by combination of a current conveyor and a voltage inverter. An independent feedback loop compensates the offset arising from asymmetries within the circuitry. We focused especially on the stability of the current source for usage with small metal electrodes in aqueous solutions. At the monitor side two identical, high input impedance difference amplifiers convert the net current through the object and the voltage dropping across into a 50 Ω symmetric output. The entire circuitry is optimized for step response making it suitable for fast time domain measurements.
[Show abstract][Hide abstract] ABSTRACT: The initial effect of nanosecond pulsed electric fields (nsPEFs) on cells is a change of charge distributions along membranes. This first response is observed as a sudden shift in the plasma transmembrane potential that is faster than can be attributed to any physiological event. These immediate, yet transient, effects are only measurable if the diagnostic is faster than the exposure, i.e., on a nanosecond time scale. In this study, we monitored changes in the plasma transmembrane potential of Jurkat cells exposed to nsPEFs of 60 ns and amplitudes from 5 to 90 kV/cm with a temporal resolution of 5 ns by means of the fast voltage-sensitive dye Annine-6. The measurements suggest the contribution of both dipole effects and asymmetric conduction currents across opposite sides of the cell to the charging. With the application of higher field strengths the membrane charges until a threshold voltage value of 1.4-1.6 V is attained at the anodic pole. This indicates when the ion exchange rates exceed charging currents, thus providing strong evidence for pore formation. Prior to reaching this threshold, the time for the charging of the membrane by conductive currents is qualitatively in agreement with accepted models of membrane charging, which predict longer charging times for lower field strengths. The comparison of the data with previous studies suggests that the sub-physiological induced ionic imbalances may trigger other intracellular signaling events leading to dramatic outcomes, such as apoptosis.
Biophysics of Structure and Mechanism 05/2011; 40(8):947-57. DOI:10.1007/s00249-011-0710-7 · 2.22 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The electrical properties of nanostructured silicon electrodes in contact with phosphate buffered saline were examined using small signal impedance together with cyclic voltammetry. The highest electrode impedance appears at thermodynamic equilibrium, i.e. at zero current potential. Biasing the electrode in both directions decreases the magnitude of the electrode impedance without showing a clear rectifier effect within ± 3V. The impact of nanostructures on the electrode impedance was negligible given the variability between single electrodes.
[Show abstract][Hide abstract] ABSTRACT: The efficiency of biogas production during anaerobic digestion depends heavily on optimal dosing ratios and stable operations which can not be achieved without accurate and reliable monitoring and control of the dry matter (DM) and organic dry matter (oDM) content. The materials in biogas processes to be measured are either stored in a vessel flowing in a pipe, either as a liquid or as solid particles in pneumatic or liquid assisted transportation with a wide area of the content of DM (1% 90). In such cases microwave sensors provide an attractive solution, because microwaves penetrate most materials allowing the non destructive measurement to be representative for a special volume or the cross section of the pipe. The setup of the measuring system for materials with high water content and first results are presented.
[Show abstract][Hide abstract] ABSTRACT: Electrical measurement is a simple innocuous tool for material characterization. Unlike for instance milk or dairy products,
the majority of naturally grown food is composed of cells. The cells of meat or vegetables are surrounded by an insulating
membrane, while the cytosol and the extracellular fluids are electrolytes. Despite the high permittivity of water, electrolytes
behave like ohmic resistors up to hundreds of MHz. In contrast, membranes form capacitive elements due to their high resistance.
The typical time constant for charging cell membranes is of the order of a microsecond. Thus, cells influence the impedance
in a frequency range up to several MHz. At higher frequencies, the cytosolic content, i.e., macromolecules, gives rise to
characteristic relaxation processes. Using the impedance in the microwave range where water dipoles show a distinct dispersion,
humidity of dried matter can be addressed. Moreover, with sensitive measurement setup and proper models, one can determine
the dry content in mashes and slurries as well. Several quality standards correlate well with the permeability of the membranes
or the total water content. Because of the comparatively simple measurement of the electrical impedance together with advanced
mathematical modeling, it is often a good approach in quality assessment of agricultural products. The change in conductivity
of a culture medium contains information about the metabolism of incubated cells. Using specific culture media and time-lapse
conductivity monitoring allows a high sensitivity and selectivity in microbial detection. Because the electrical impedance
is very sensitive to the permeability of cell membranes, it is a great choice for the assessment of changes due to high voltage
application. Today, many attempts to use impedance measurement in food technology show fast success in research but fail in
practice. The reason is often an overestimation especially of the selectivity while underestimating the uncertainties in a
harsh environment of a food-processing plant. Established, however, is the use of robust process measurement systems and the
limitation of impedance measurement to applications with highly significant outcome or as supplemental measurement in a multiparameter
approach. This review introduces the basics of bioimpedance measurement, points to sources of uncertainty, and presents successful
applications in food industry.
KeywordsBioimpedance-Food quality-Drip loss-Microbial detection-Water content