Impedance analysis of epithelial and endothelial cell monolayers cultured on gold surfaces

Wegener, Joachim; Sieber, Manfred; Galla, Hans‐Joachim

doi:10.1016/0165-022x(96)00005-x

Cited by 121 publications

(100 citation statements)

References 24 publications

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“…8 shows fluorescence microscopic images of immuno-stainings directed against zonula occludens protein 1, the first protein that was identified to be a unique constituent of the tight junctions. ZO-1 is an intracellular peripheral membrane protein that serves as a linker protein between the actin cytoskeleton and the junction forming transmembrane proteins (Wegener et al, 1996). We prepared samples of PBCEC monolayers that were cultured in serum-free medium without HC addition (upper panel) or in presence of HC (lower panel).…”

Section: Resultsmentioning

confidence: 99%

“…Impedance contributions arising from the cell-substrate adhesion zone are accounted for by the parameter ␣ that is defined as Top and side view of the completed measurement chamber with the electrode holding slide as the chamber bottom. For details of chamber construction see (Wegener et al, 1996). During data recording chambers were placed in a humidified incubator (37 • C).…”

Section: Electric Cell-substrate Impedance Sensing (Ecis)mentioning

confidence: 99%

“…Thus, ECIS data recorded for the three working electrodes provide independent readings from three individual spots within the cell layer. The electrode arrangement is prepared by high-vacuum thermal evaporation of gold through a corresponding mask onto an ordinary microscope slide as described elsewhere (Wegener et al, 1996). A glass top with a screw cap is mounted onto the slide by a non-cytotoxic silicon glue forming a chamber that can be handled like a common cell culture dish (Fig.…”

Section: Electric Cell-substrate Impedance Sensing (Ecis)mentioning

confidence: 99%

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Bioelectrical impedance assay to monitor changes in cell shape during apoptosis

Arndt

Seebach

Psathaki

et al. 2004

Biosensors and Bioelectronics

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299

197

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Apoptosis is a strictly regulated and genetically encoded cell 'suicide' that may be triggered by cytokines, depletion of growth factors or certain chemicals. It is morphologically characterized by severe alterations in cell shape like cell shrinkage and disintegration of cell-cell contacts. We applied a non-invasive electrochemical technique referred to as electric cell-substrate impedance sensing (ECIS) in order to monitor the apoptosis-induced changes in cell shape in an integral and quantitative fashion with a time resolution in the order of minutes. In ECIS the cells are grown directly on the surface of small gold-film electrodes (d = 2 mm). From readings of the electrical impedance of the cell-covered electrode, performed with non-invasive, low amplitude sensing voltages, it is possible to deduce alterations in cell-cell and cell-substrate contacts. To improve the sensitivity of this impedance assay we used endothelial cells derived from cerebral micro-vessels as cellular model systems since these are well known to express electrically tight intercellular junctions. Apoptosis was induced by cycloheximide (CHX) and verified by biochemical and cytological assays. The time course of cell shape changes was followed with unprecedented time resolution by impedance readings at 1 kHz and correlated with biochemical parameters. From impedance readings along a broad frequency range of 1-10 6 Hz we could assign the observed impedance changes to alterations on the subcellular level. We observed that disassembly of barrier-forming tight junctions precedes changes in cell-substrate contacts and correlates strongly with the time course of protease activation.

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Section: Resultsmentioning

confidence: 99%

Section: Electric Cell-substrate Impedance Sensing (Ecis)mentioning

confidence: 99%

Section: Electric Cell-substrate Impedance Sensing (Ecis)mentioning

confidence: 99%

See 1 more Smart Citation

Bioelectrical impedance assay to monitor changes in cell shape during apoptosis

Arndt

Seebach

Psathaki

et al. 2004

Biosensors and Bioelectronics

Self Cite

299

197

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“…Most of the work in this field has centered on the use of frequencies in the radio-wave range from 10 Hz to 100 MHz. [7][8][9] The principal thrust of this prior work has been to elucidate the basic dielectric properties of intact cells and cell membranes in an attempt to observe cellular behavior in a noninvasive fashion. [7][8][9] In addition to these efforts, some researchers have attempted to monitor cellular ligand-receptor interactions using bioimpedance.…”

Section: Introductionmentioning

confidence: 99%

“…[1][2][3][4][5][6][7][8][9][10] The technology employs the application of alternating voltages at set frequencies to cellular components (membranes) and intact cells and monitors the resulting currents generated from the dipoles within the sample. At each frequency, the difference in magnitude of the reflected signal, as compared to the incident wave, is measured.…”

Section: Introductionmentioning

confidence: 99%

Cellular Dielectric Spectroscopy: A Powerful New Approach to Label-Free Cellular Analysis

Ciambrone¹,

Liu²,

Lin³

et al. 2004

SLAS Discovery

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The past decade has seen a number of significant changes in identifying higher quality lead compounds earlier in the drug discovery process. Cell-based assay technologies yielding high-content information have emerged to achieve this goal. Although most of these systems are based on fluorescence detection, this article describes the development and application of an innovative cellular assay technology based on radio frequency spectrometry and bioimpedance measurements. Using this technique, the authors have discovered a link between cellular bioimpedance changes and receptor-mediated signal transduction events. By performing dielectric spectroscopy of cells across a spectrum of frequencies (1 KHz to 110 MHz), a series of receptorspecific, frequency-dependent impedance patterns is collected. These raw data patterns are used to determine the identity of the cellular receptor-signaling pathway being tested and to quantify stimulation endpoints and kinetics. The authors describe the application of this technology to the analysis of ligand-induced cellular responses mediated by the 3 major classes of Gprotein-coupled receptors (GPCRs) and protein tyrosine kinase receptors. This single assay platform can be used with ease to monitor G s , G i , and G q GPCRs without the need for chimeric or promiscuous G-proteins, fluorophors, or tagged proteins. In contrast to other methods of monitoring cellular signal transduction, this approach provides high information content in a sim-

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Porcinechoroid plexus epithelial cells in culture: Regulation of barrier properties and transport processes

Haselbach

Wegener

Decker

et al. 2000

Microsc. Res. Tech.

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The epithelial cells of the choroid plexus are the structural basis of the blood‐cerebrospinal fluid (CSF)‐barrier. Here we summarise our recent efforts to culture those cells mainly on permeable supports in vitro. Isolated from porcine brains, we report a simple protocol for the primary culture using cytosine arabinoside as an additive that is cytotoxic for other cells except the plexus epithelial cells. Enhanced barrier properties are obtained by withdrawal of serum from the culture medium after confluency is reached. Cells improve their polarity, permeability for hydrophilic substrates is lowered, electrical resistance is increased tenfold, and a pH‐gradient is built up across the cell monolayer. Polarised secretion of proteins and most importantly fluid secretion into the apical filter compartment was attained and proven to be dependent on the Na+,K+‐ATPase activity. Active transport processes (penicillin G, riboflavin, myo‐inositol, ascorbic acid) were studied and clearly showed the involvement of the organic anion transporter. The permeability of the barrier was found to be regulated by cyclic adenosine monophosphate (cAMP). Moreover, we report that cell proliferation and differentiation is controlled by components of the extracellular matrix. The present culture model could now be used as an in vitro system to quantify drug transport across the blood‐CSF‐barrier. Microsc. Res. Tech. 52:137–152, 2001. © 2001 Wiley‐Liss, Inc.

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Impedance analysis of epithelial and endothelial cell monolayers cultured on gold surfaces

Cited by 121 publications

References 24 publications

Bioelectrical impedance assay to monitor changes in cell shape during apoptosis

Bioelectrical impedance assay to monitor changes in cell shape during apoptosis

Cellular Dielectric Spectroscopy: A Powerful New Approach to Label-Free Cellular Analysis

Porcinechoroid plexus epithelial cells in culture: Regulation of barrier properties and transport processes

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