Impedance analysis of adherent cells after in situ electroporation: Non-invasive monitoring during intracellular manipulations

Stolwijk, Judith A.; Hartmann, Christoph; Balani, Poonam; Albermann, Silke; Keese, Charles R.; Giæver, Ivar; Wegener, Joachim

doi:10.1016/j.bios.2011.05.033

Cited by 58 publications

(59 citation statements)

References 36 publications

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“…4B). Our apoptotic data and previous experiments correlated such changes to the loss of cellular monolayer resulting from cell death and cell de-attachment (Arndt et al, 2004; Stolwijk et al, 2011). Contrary, cells exposed to 10 and 25 nM digitoxin showed a significant increase in α relative to controls, indicating a stimulation of their cellular attachment.…”

Section: Resultssupporting

confidence: 85%

Real-time analysis of the effects of toxic, therapeutic and sub-therapeutic concentrations of digitoxin on lung cancer cells

Eldawud

Stueckle

Manivannan

et al. 2014

Biosensors and Bioelectronics

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Digitoxin belongs to a naturally occurring class of cardiac glycosides (CG); digitoxin is clinically approved for heart failure and known for its anti-cancer effects against non-small lung cancer cells (NSCLC). However, concerns associated with its narrow therapeutic index and its concentration-dependent mechanism of action are rising. Thus, before digitoxin implementation in designing and developing safer and more effective CG-based anti-cancer therapies, its pharmacological and safety profiles need to be fully elucidated. In this research we used a combinatorial approach to evaluate the anti-cancer mechanisms of digitoxin in real-time. Our approach employed a non-invasive electric cell impedance sensing technique as a proxy to monitor NSCLC behavior post-exposure to toxic, therapeutic and sub-therapeutic concentrations of the drug. By developing structure–function combinatorial relations we showed that digitoxin targets cancer cells in a time and dose-dependant manner by activating pro-apoptotic and anti-proliferative signaling cascades that results in strengthening cellular adhesion and sequestration of key regulatory proliferation protein from the nucleus.

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Section: Resultssupporting

confidence: 85%

Real-time analysis of the effects of toxic, therapeutic and sub-therapeutic concentrations of digitoxin on lung cancer cells

Eldawud

Stueckle

Manivannan

et al. 2014

Biosensors and Bioelectronics

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“…This may be because, as previously stated, for 800 V/cm and 1200 V/cm irreversible damage is produced in the cell membrane. The obtained values are in accordance with observations made previously by other authors who also measured durations of around one second for the total lifetime of the short-term membrane resealing mechanisms [21,[54][55][56][57].…”

Section: Accepted M Manuscriptsupporting

confidence: 81%

“…Electrical measurements have usually been performed by observing the changes in the applied current-voltage waveform, which mainly consist of quasi-dc conductivity changes. Other attempts have used AC signals to study the passive electrical properties of the sample before and after electroporation at single frequencies [21][22][23]. Only in a few studies have multifrequency measurements been performed, also referred to as electrical impedance spectroscopy (EIS), on electroporated samples [20,24,25].…”

Section: Accepted M Manuscriptmentioning

confidence: 99%

Interpulse multifrequency electrical impedance measurements during electroporation of adherent differentiated myotubes

García-Sánchez

Azan

Leray

et al. 2015

Bioelectrochemistry

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In this study, electrical impedance spectroscopy measurements are performed during electroporation of monolayers of differentiated myotubes. The time resolution of the system (1 spectrum/ms) enable 860 full spectra (21 frequencies from 5 kHz to 1.3 MHz) to be acquired during the time gap between consecutive pulses (interpulse) of a classical electroporation treatment (8 pulses, 100 µs, 1 Hz). Additionally, the characteristics of the custom microelectrode assembly used allow the experiments to be performed directly in situ in standard 24 multi-well plates. The impedance response dynamics are studied for three different electric field intensities (400, 800 and 1200 V/cm).The multifrequency information, analysed with the Cole model, reveals a short-term impedance recovery after each pulse in accordance with the fast resealing of the cell membrane, and a longterm impedance decay over the complete treatment in accordance with an accumulated effect pulse after pulse. The analysis shows differences between the lowest electric field condition and the other two, suggesting that different mechanisms that may be related with the reversibility of the process are activated. As a result of the multifrequency information, the system is able to measure simultaneously the conductivity variations due to ion diffusion during electroporation.Finally, in order to reinforce the physical interpretation of the results, a complementary electrical equivalent model is used.

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“…Coplanar patterning of thin-film electrodes on cell culture ware makes solid substrate-based impedance measurements consistent and compatible with in vitro cell culture, immunofluorescence staining and biochemical assays. Even combinations with other electrical methods that use thin-film electrodes can be easily achieved, such as in situ electroporation of cells [95], Quartz Crystal Microbalance (QCM) [44], Surface Plasmon Resonance (SPR) [68], that allow for multiparametric analysis of the cell culture. Substrate-integrated electrodes can either be arranged as interdigitated comb structures (IDEs) [31,71,92,5] or use a small working electrode (WE) in combination with a large counter electrode (CE) (Fig.…”

Section: Overview Of Endothelial Barrier Function Assaysmentioning

confidence: 99%

“…In a cell layer with a transfection efficiency of 80% it is still possible that none of the cells on the actual electrode is actually transfected (extreme case, 8W1E). In situ electroporation using the ECIS electrodes to introduce siRNA, cDNA or other macromolecules into cells [95] could provide an interesting possible solution, as transfection is restricted to the cell population directly on the electrode. Toxic side effects of transfection reagents or massive protein overexpression can significantly affect survival rate and the process of cell layer maturation, and as a consequence baseline resistance of the cell layer.…”

Section: Experimental Settings and Practical Guidelinesmentioning

confidence: 99%

Impedance analysis of GPCR-mediated changes in endothelial barrier function: overview and fundamental considerations for stable and reproducible measurements

Stolwijk

Matrougui

Renken

et al. 2014

Pflugers Arch - Eur J Physiol

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101

116

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The past 20 years have seen significant growth in using impedance-based assays to understand the molecular underpinning of endothelial and epithelial barrier function in response to physiological agonists, pharmacological and toxicological compounds. Most studies on barrier function use G protein coupled receptor (GPCR) agonists which couple to fast and transient changes in barrier properties. The power of impedance based techniques such as Electric Cell-Substrate Impedance Sensing (ECIS) reside in its ability to detect minute changes in cell layer integrity label-free and in real-time ranging from seconds to days. We provide a comprehensive overview of the biophysical principles, applications and recent developments in impedance-based methodologies. Despite extensive application of impedance analysis in endothelial barrier research little attention has been paid to data analysis and critical experimental variables, which are both essential for signal stability and reproducibility. We describe the rationale behind common ECIS data presentation and interpretation and illustrate practical guidelines to improve signal intensity by adapting technical parameters such as electrode layout, monitoring frequency or parameter (resistance versus impedance magnitude). Moreover, we discuss the impact of experimental parameters, including cell source, liquid handling and agonist preparation on signal intensity and kinetics. Our discussions are supported by experimental data obtained from human microvascular endothelial cells challenged with three GPCR agonists, thrombin, histamine and Sphingosine-1-Phosphate.

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Impedance analysis of adherent cells after in situ electroporation: Non-invasive monitoring during intracellular manipulations

Cited by 58 publications

References 36 publications

Real-time analysis of the effects of toxic, therapeutic and sub-therapeutic concentrations of digitoxin on lung cancer cells

Real-time analysis of the effects of toxic, therapeutic and sub-therapeutic concentrations of digitoxin on lung cancer cells

Interpulse multifrequency electrical impedance measurements during electroporation of adherent differentiated myotubes

Impedance analysis of GPCR-mediated changes in endothelial barrier function: overview and fundamental considerations for stable and reproducible measurements

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