Characterization of antimicrobial peptide activity by electrochemical impedance spectroscopy

Electrochemical impedance spectroscopy performed on surface-supported bilayer membranes allows for the monitoring of changes in membrane properties, such as thickness, ion permeability, and homogeneity, after exposure to antimicrobial peptides (AMPs). We show that two model cationic peptides, very similar in sequence but different in activity, induce dramatically different changes in membrane properties as probed by impedance spectroscopy. Moreover, the impedance results excluded the “barrel-stave” and the “toroidal pore” models of AMP mode of action, and are more consistent with the “carpet” and the “detergent” models. The impedance data provide important new insights about the kinetics and the scale of the peptide action which currently are not addressed by the “carpet” and the “detergent” models. The method presented not only provides additional information about the mode of action of a particular AMP, but offers a means of characterizing AMP activity in reproducible, well-defined quantitative terms.     

The electrical impedance spectroscopy of Scots pine needles during cold acclimation

The electrical impedance spectroscopy (EIS) was applied to current-year needles of Scots pine ( Pinus sylvestris L.) in an 8-year provenance field trial in central Finland during frost hardening. The EIS analysis of the needles using a Model-A equivalent circuit indicated a sequence of events in the needles during their cold acclimation. Some of the EIS-parameters referred to maturation phenomena occurring during the pre-hardening phase at the end of the growing season, and some parameters displayed a clear coincidence with the frost hardening itself. Significant differences between provenances were found in several of the Model-A parameters. Extracellular resistance (r_e) and β -coefficient decreased in all provenances in the pre-hardening phase in August and until mid-September. In the same phase, both the intracellular resistance (r_i) and the cell membrane time constant (τ_m) first increased and then decreased. According to τ_m, r_e and β there was a clear gradation between provenances in the pre-hardening phase. From the end of September significant differences were found in the intracellular resistance between provenances, corresponding with the differences in their hardening pattern. The dry weight (DW) content of needles increased during the study period but no clear differences were found between the provenances.     

Electrical characterization of phytoplankton suspensions using impedance spectroscopy

Journal of Applied Phycology - This study used impedance spectroscopy measurements to extract the electrical properties of phytoplankton cells in suspension. Experimental measurements were...     

Detection of phenolic compounds using impedance spectroscopy measurements

Langmuir-Blodgett (LB) and layer-by-layer films (LbL) of a PPV (p-phenylenevinylene) derivative, an azo compound and tetrasulfonated phthalocyanines were successfully employed as transducers in an “electronic tongue” system for detecting trace levels of phenolic compounds in water. The choice of the materials was based on their distinct electrical natures, which enabled the array to establish a fingerprint of very similar liquids. Impedance spectroscopy measurements were taken in the frequency range from 10 Hz to 1 MHz, with the data analysed with principal component analysis (PCA). The sensing units were obtained from five-layer LB films of (poly[(2-methoxy-5-n-hexyloxy)-p-phenylenevinylene]), OC₁OC₁₈-PPV (poly(2-methoxy,5-(n-octadecyl)-p-phenylenevinylene)), DR (HEMA-co-DR13MA (poly-(hydroxyethylmethacrylate-co-[4′-[[2-(methacryloyloxy)-ethyl]ethylamino]-2-chloro-4-nitroazobenzene]))) and five-bilayer LbL films of tetrasulfonated metallic phthalocyanines deposited onto gold interdigitated electrodes. The sensors were immersed into phenol, 2-chloro-4-methoxyphenol, 2-chlorophenol and 3-chlorophenol (isomers) solutions at 1 × 10⁻⁹ mol L⁻¹, with control experiments carried out in ultra pure water. Samples could be distinguished if the principal component analysis (PCA) plots were made with capacitance values taken at 10³ Hz, which is promising for detection of trace amounts of phenolic pollutants in natural water.     

Correlation of electrical impedance and temperature in tissue during freezing.

Experiments were performed correlating tissue temperature and tissue impedance in the course of freezing canine skin and palate. Tissue impedance rose gradually to high levels as the tissue froze. Tissue temperature of ?26 ° to ?30 °C corresponded closely to tissue impedance of 1 MΩ. Tissue impedance of 5 to 10 MΩ corresponded closely to temperatures of ?40 °C and colder. These observations indicated that the measurement of tissue impedance was related to tissue temperature and suggested that either or both could be used to predict tissue destruction in cryosurgery.     

Electrical impedance spectroscopy measurements using a four-electrode configuration improve on-line monitoring of cell concentration in adherent animal cell cultures

This paper describes the improvement in the use of electrical impedance spectroscopy (EIS) for animal cell concentration monitoring of adherent cultures by using a four-electrode configuration instead of the commonly used two-electrode configuration. This four-electrode configuration prevents cell concentration measurements from external masking effects such as the electrode covering ratio, the degree of cellular adherence to the electrodes and the impedance of the measuring electrodes. Cell concentration was monitored using both four-electrode and two-electrode configurations in vero cell and human mesenchymal stem cell cultures in order to analyze the attained improvement in two cell lines with opposite growth characteristics. The experiments performed with vero cell cultures evidenced that the four-electrode configuration enables cell concentration measurements along all culture phases, even once the culture reached cell confluence (over 2×10(5) cells/cm(2)), confirming that this configuration is less effected by all the external influences. The experiments performed with human mesenchymal stem cells demonstrated good sensitivity of the measurement at very low cell concentrations, as well as a very good robustness all over the 12-days experiment. Finally, off-line cell measurements during cell cultures proved good accuracy of impedance measurements carried out with a four-electrode configuration along all cell growth phases, enabling determination of relevant cell growth parameters.     

Magnetic field influence on electrical properties of human blood measured by impedance spectroscopy

The impedance spectroscopy technique (IST) was used for studying the effect of a 0.5 T magnetic field on the electrical properties of whole human blood. A Solartron SI 1260 spectrometer was used to measure the impedance spectra of magnetic field exposed blood samples compared to non-exposed samples. An equivalent electrical circuit model, consisting in a resistance Rs in series with a parallel circuit formed by a constant phase element (CPE) and another resistance Rp, is proposed to fit the data in both cases. The experiment used 3 ml human blood samples from 160 healthy donors. A Wilcoxon matched pairs statistical test was applied to the data. The data analysis seems to show a statistically significant increase of the values of resistance Rp (Z = 5.06, P < 0.001) and capacitance CT (Z = 3.32, P < 0.001) of the blood exposed to magnetic field, by approximately 10.4% and 1.9%, respectively.     

On-line monitoring of lipid storage in yeasts using impedance spectroscopy

Bioremediation technologies and many environmentally sound biosyntheses rely on the catalytic potential of whole cells. For analyzing and controlling such processes robust real-time indicators for the concentration of intact cells such as impedance are required. The conventional method measures the capacitances of cell suspensions at one or two frequencies and correlates them with biomass concentrations. However, cell inclusions such as lipid droplets or overproduced enzymes may block intracellular ion paths, thereby possibly modifying the dielectric properties of the cells. To test the hypothesis that the total impedance spectrum into the analysis may provide useful information about cell inclusions, the impedance spectrum of a technical culture of the oleaginous yeast Arxula adeninivorans was measured and evaluated every 15 s. This yeast is a good test object since it stores the excess of assimilated carbon in experimentally controllable lipid droplets. Upon correction for possible impedance signal interferences, we derived different empirical methods suitable to indicate incipient lipid formation. The methods were designed to act on-line and are thus principally suited for real-time monitoring of cell inclusions. In search for optimised bioprocess monitoring we tested a heuristic spectrum analysis using integrative statistics (RDA). With this approach we were able to accurately detect the formation of cell inclusions, which is potentially valuable for future bioprocess control strategies.     

In vitro electrical impedance spectroscopy of human dentine: the effect of restorative materials

The influence of different restorative materials on in vitro dielectric properties of sound dentine was investigated. The studied samples were three-layer materials consisting of successive disks of dentine and silver amalgam or nanohybrid composite resin. Before being tested, the samples were maintained in physiological solution never more than 48 h from the extraction. Also, sections of intact dentine were similarly prepared for electrical measurements. Complex dielectric permittivity of these specimens was determined in a wide frequency range using the parallel-plate capacitor technique. Very similar dielectric responses of intact dentine and amalgam-dentine material were observed. This is explained on the basis of high dc conductivity exhibited by both samples. In contrast, resin-dentine specimen revealed a much more insulating behavior. A simple theoretical model for heterogeneous systems could be applied to these dental three-layer materials. The dielectric properties of restored dentine are strongly dependent on the kind of restorative material employed in each case. This suggests that electrical data should be used carefully in caries diagnosis on restored teeth.     

Characterization of bacteria contaminating tissue cultures of apple rootstock 'YP'

Meristem-tip cultures of apple rootstock 'YP' were started at different times of the year over a period of 2 years and the contamination of the cultures was monitored during five subcultures. Bacterial contaminants were isolated to pure cultures, identified by the API test system and appropriate additional tests, and the sensitivity of the most common isolates to different antibiotics was determined. Of the 216 strains isolated from the initiation cultures, 78% were pseudomonads, coryneforms or enterobacteria. Only three bacterial contaminants were found at the multiplication stage. A greater part of the contaminants were likely to originate from the stock plant. Rifampicin (at 50–200 mg 1^-1) and cefotaxime (at 250–1500 mg 1^-1) were found to be bactericidal against many isolates, but differences between species and strains were found.     

Combining optical and electrical impedance techniques for quantitative measurement of confluence in MDCK-I cell cultures

A new method combining optical and electrical impedance measurements is described that enables submicroscopic cell movements to be monitored. The cells are grown on small gold electrodes that are transparent to light. This modified electrical cell-substrate impedance sensor (ECIS) allows simultaneous microscopic recording of both growth and motility, thus enabling cell confluence on the electrodes to be systematically correlated to the impedance in regular time intervals of seconds and for extended periods of time. Furthermore, the technique provides an independent measure of monolayer cell densities that we compare to calculated values from a theoretical model. We have followed the attachment and spreading behavior of epithelial Madin-Darby canine kidney strain I (MDCK-I) cell cultures on microelectrodes for up to 40 h. The studies reveal a high degree of correlation between the measured resistance at 4 kHz and the corresponding cell confluence in 4- to 6-h intervals with typical linear cross-correlation factors of r equaling approximately 0.9. In summary, the impedance measured with the ECIS technique provides a good quantitative measure of cell confluence.     

Some early results related to electrical impedance of normal and abnormal gastric tissue

Gastric cancer is the fourth most common cancer and most patients with gastric cancer are being diagnosed in advanced stages of the disease so they do not gain any survival chance from conventional surgical, chemotherapeutic or radiotherapeutic methods. These are relatively high cost procedures in terms of both time and money. This study considers the introduction of a novel minimally invasive diagnostic technique which shows the relationship between histopathology and the electrical impedance spectrum in the human stomach. In this study, 4 electrode technique was used to differentiate tissues from each other using Tabriz Mark 1 electrical impedance system (30 different frequencies in the range of 2 kHz to 1 MHz). A total of 97 points from 45 patients were studied in terms of their biopsy reports matching to the electrical impedance measurements (in vivo). After impedance measurements and applying calibration factors, a non-parametric statistical technique, the Kruskal–Wallis test was used to evaluate the difference among the groups. According to the calculation of respective data using this spectroscopy system, the resistivity of the normal group was higher than that of the benign group, and the resistivity of these groups were higher than that of the malignant group at frequencies between 470 kHz and 1 MHz (P < 0.05). In these frequencies, the impedivity of the dysplastic tissue was significantly lower than that of the other groups (P < 0.05). Also, Cole equation fitting procedure was used to generate a scatter plot of the malignant and benign points: it shows in general, benign points had higher values of R than the malignant points. Therefore, electrical impedance spectroscopy can be a useful technique to characterize the stomach tissue.     

In-situ biomass characterisation by impedance spectroscopy using a full-bridge circuit

A full-bridge sensor system for characterising biosuspensions by impedance spectroscopy which avoids cross-sensitivity to temperature is presented. To record a large range of different kinds of cells, frequency is varied between 500 kHz and 50 MHz. The characteristic frequencies of the #-polarisation of animal cells (mouse), yeasts (Saccharomyces cerevisiae) and Escherichia coli bacteria are located in this range. The widespread applicability of this method is demonstrated by measurements on suspensions of different cells and the determination of concentration in a fermentation process. Furthermore, yeast suspensions are characterised at temperatures between T₁=24°C and T₂=36°C to evaluate the influence of temperature. A reduction of cross-sensitivity of about one order of magnitude compared to a single-sensor results. Changing in the conductivity of the suspension by 26% alters the measured biomass concentration by only 2.8%.     

In vivo electrical impedance spectroscopic monitoring of the progression of radiation-induced tissue injury.

This study evaluates the potential of electrical impedance spectroscopy (EIS) as a noninvasive technique for tracking the progression of radiation-induced damage in normal muscle tissue. Male Sprague-Dawley rats were irradiated locally to the gastrocnemius and biceps femoris muscle. Single doses were administered using a procedure that spares skin and bone. Complex impedance spectral measurements (taken at 50 frequency points between 1 kHz and 1 MHz) were made at monthly intervals using recessed disk electrodes applied to the skin. A histological scoring scheme was developed for evaluation of injury. A strong dose-dependent progression of injury evident in both spectral measurements and histological scoring has been observed. Latent time also appears to be dependent on dose with changes induced by 70 Gy evident by 2 months, changes induced by 90 Gy observed by 1 month, and dramatic changes found within 3 weeks at 150 Gy. Injury was morphologically comparable to the type of damage that occurs in response to small, fractionated doses, but on a much shorter time scale. Increased spectral shift was a consistent indicator of the extent of tissue injury at the time of measurement. The use of a large single dose resulted in an excellent model in terms of inducing a significant progression in tissue injury over a short post-treatment follow-up period in the muscle mass while also providing a consistent location for in vivo electrical impedance measurements. The results show that EIS can follow radiation-induced tissue change, suggesting that EIS has the potential to monitor the types of injury observed in late radiation damage of muscle tissue noninvasively.