Sciospec - Electrical Impedance. At its best.
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Sciospec Scientific Instruments GmbH
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Deep brain stimulation (DBS) is effective for the treatment of patients with Parkinson’s disease (PD), especially in advanced stages which are refractory to conventional therapy. Despite of the regular use in clinical therapy, rodent models for basic research into DBS are not routinely available. The main reason is the geometry difference from rodents to humans, imposing larger problems in the transfer of the stimulation conditions than from primates to humans. For rodents, the development of miniaturized mobile stimulators and stimulation parameters, as well as improved electrode materials and geometry are desirable. The impedance of custom made, cylindrical (contact diameter 200 µm, length 100 µm), platinum/iridium electrodes has been measured in vivo for two weeks to characterize the influence of electrochemical processes and of the adherent cell growth at the electrode surface. During the encapsulation process, the real
part of the electrode impedance at 10 kHz doubled with respect to its initial value after a characteristic decrease by approximately one third at the second day. An outlook is given on further investigations with different electrode designs for long-term DBS.
Deep brain stimulation (DBS) is an invasive therapeutic option for patients with Parkinson’s disease (PD) but the mechanisms behind it are not yet fully understood. Animal models are essential for basic DBS research, because cell based in-vitro techniques are not complex enough. However, the geometry difference between rodents and humans implicates transfer problems of the stimulation conditions. For rodents, the development of miniaturized mobile stimulators and adapted electrodes are desirable. We implanted uni- and bipolar platinum/iridium electrodes in rats and were able to establish chronical instrumentation of freely moving rats (3 weeks). We measured the impedance of unipolar electrodes in-vivo to characterize the influence of electrochemical processes at the electrode-tissue interface. During the encapsulation process, the real part of the electrode impedance at 10 kHz doubled after 12 days and increased almost 10 times after 22 days. An outlook is given on the quantification of the DBS effect by sensorimotor behavioral tests
Unexpected adverse effects on the cardiovascular system remain a major challenge in the development of novel active pharmaceutical ingredients (API). To overcome the current limitations of animal-based in vitro and in vivo test systems, stem cell derived human cardiomyocyte clusters (hCMC) offer the opportunity for highly predictable pre-clinical testing. The three-dimensional structure of hCMC appears more representative of tissue milieu than traditional monolayer cell culture. However, there is a lack of long-term, real time monitoring systems for tissue-like cardiac material. To address this issue, we have developed a microcavity array (MCA)-based label-free monitoring system that eliminates the need for critical hCMC adhesion and outgrowth steps. In contrast, feasible field potential derived action potential recording is possible immediately after positioning within the microcavity. Moreover, this approach allows extended observation of adverse effects on hCMC. For the first time, we describe herein the monitoring of hCMC over 35 days while preserving the hCMC structure and electrophysiological characteristics. Furthermore, we demonstrated the sensitive detection and quantification of adverse API effects using E4031, doxorubicin, and noradrenaline directly on unaltered 3D cultures. The MCA system provides multi-parameter analysis capabilities incorporating field potential recording, impedance spectroscopy, and optical read-outs on individual clusters giving a comprehensive insight into induced cellular alterations within a complex cardiac culture over days or even weeks.
Wound infection status is a relevant diagnostic parameter to enhance wound treatment towards better healing rate. Impedance evaluation is a powerful tool to measure the inflammatory response like the released DNA of neutrophils. In our research we investigated the dielectric behaviour of neutrophils settled on electrodes in vitro. The cells have been stimulated to react in the same way as in a wound infection. The result is a significant impedance deviation of about 50 % with comparable amount of cells like in an infected wound. Microscopic fluorescence verifications acknowledge these findings.
Wound infection monitoring is a challenging task. It is only solvable by designing an integrable and cost-efficient sensor which measures a relevant set of parameters. One viable parameter is the formation of neutrophil extracellular traps (NETs). Their task is trapping pathogens in the wound. A wound infection results in massive release of them which can be detected with impedimetric methods. Our investigations focused on the characterization of the biological process with an in vitro model. The model environment is a cell culture with neutrophil granulocytes cultured on interdigitated electrodes which represent the sensor surface. Detected impedance changes caused by NET-formation were in the range of 35% and even higher. This implies that impedance measurements are suitable for NET detection. We derived a measurement and evaluated it by differing conditions like changing stimulation agent and varying the cell number. For both conditions the results of impedance and phase angle deviation can be confirmed. In combination with other parameters a sensor can be designed for specific detection of wound infections. These aspects are integrated in our sensor concept.
Biofilm formation can cause serious health hazards, mostly due to the uncontrolled release of pathogens. This can generate several problems in industrial facilities, e.g., in the food industry. The aim of the present study was to develop and implement a multi-parametric sensor system to monitor biofilm formation in laboratory as well as industrial set-ups. To minimize cross sensitivity or interference, the device was based on a combination of different measurement principles. Micro-organisms were initially cultivated in a laboratory scale reactor. Afterwards, biofilm formation will be studied with each prototype of the multi-parametric sensor followed by final tests on an industrial scale.
Aggressive cancer entities like neuroblastoma and glioblastoma multiforme are still difficult to treat and have discouraging prognosis in malignant stage. Since each tumor has its own characteristics concerning the sensitivity towards different chemotherapeutics and moreover, can obtain resistance, the development of novel chemotherapeutics with a broad activity spectrum, high efficacy and minimum side effects is a continuous process. Sophisticated in vitro assays for comprehensive prediction of in vivo drug efficacy and side effects represent an actual bottleneck in the drug development process. In this context, we developed a novel in vitro 2D and 3D multiwell–multielectrode device for drug efficacy monitoring based on direct real-time impedance spectroscopy measurement in combination with our unique 96-well multielectrode arrays and microcavity arrays. For demonstration, we used three neuro- and glioblastoma cell lines that were cultured as monolayer and multicellular tumor spheroids for recapitulating in vivo conditions. Using our novel 96-well multielectrode array based system it was possible to detect time and concentration dependent responses concerning treatment with doxorubicin, etoposide and vincristine. While all tested chemotherapeutics revealed high potency for apoptosis induction in neuroblastoma cells, etoposide was ineffective for glioblastoma cell lines. Determination of IC50 values allowed us to compare drug efficacy in 2D and 3D culture models and moreover, revealed chemotherapeutic and tumor cell line specific activity patterns. These pharmacokinetic patterns are of great interest in the context of preclinical drug development. Thus, impedance spectroscopy based monitoring systems could be used for the fast in vitro based in vivo prediction of novel anti-tumor drugs.
Label-free and non-invasive monitoring of receptor activation and identification of the involved signal pathways in living cells is an ongoing analytic challenge and a great opportunity for biosensoric systems. In this context, we developed an impedance spectroscopy-based system for the activation monitoring of NPY-receptors in living cells. Using an optimized interdigital electrode array for sensitive detection of cellular alterations, we were able for the first time to quantitatively detect the NPY-receptor activation directly without a secondary or enhancer reaction like cAMP-stimulation by forskolin. More strikingly, we could show that the impedimetric based NPY-receptor activation monitoring is not restricted to the Y1-receptor but also possible for the Y2- and Y5-receptor. Furthermore, we could monitor the NPY-receptor activation in different cell lines that natively express NPY-receptors and proof the specificity of the observed impedimetric effect by agonist/antagonist studies in recombinant NPY-receptor expressing cell lines. To clarify the nature of the observed impedimetric effect we performed an equivalent circuit analysis as well as analyzed the role of cell morphology and receptor internalization. Finally, an antagonist based extensive molecular signal pathway analysis revealed small alterations of the actin cytoskeleton as well as the inhibition of at least L-type calcium channels as major reasons for the observed NPY-induced impedance increase. Taken together, our novel impedance spectroscopy based NPY-receptor activation monitoring system offers the opportunity to identify signal pathways as well as for novel versatile agonist/antagonist screening systems for identification of novel therapeutics in the field of obesity and cancer.
Sensors for monitoring wound infections are important to improve care management especially for chronic wounds. As detection parameter the formation of extracellular chromatin was chosen which has characteristic dielectric properties in ionic solvents due to its bound negative charges. Experiments with planar electrodes resulted in a high impedance increase of nearly 450%. The analysis of the relative permittivity revealed a cut-off frequency at 5 kHz. It is shown for the very first time that the changing electrical medium properties during Neutrophil Extracellular Traps (NET) formation are relevant for the occurring dispersion. A textile sensor set-up is proposed to fulfill the requirements of miniaturization and bio-compatibility. With these experimental results it is possible to design a fiber-based sensor based on an impedance detection principle.
The so-called NETs (neutrophil extracellular traps) are found in vast amounts in inflamed wounds and are therefore a good marker for wound infections. They are a product of an immune reaction. Their integrant DNA has a certain dielectric behavior due to its charge. This allows a direct electric determination without the need of a transducer. Human neutrophils were used to measure the release of NETs in vitro. However, the structural changes of the cells during this process have to be taken into account. In this work a model was developed which reflects these changes. This model was compared with impedance measurements. We found that changes in the medium composition strongly modify the dielectric behavior of the system. The most obvious change here is caused by the appearance of the NETs. These changes remain also stable after the cells died and did not undergo more structural changes. The measurement of NETs is a very promising approach to support the diagnosis of inflammation processes especially in wounds.