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Ecis zθ system

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The ECIS Zθ system is an instrument used for real-time monitoring and analysis of cellular behaviors and interactions. It measures the electrical properties of cells in culture, providing insights into their physiological state, attachment, and responses to various stimuli.

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Lab products found in correlation

Collagen 4 coated gold electroarray 96 well plates, by Applied Biophysics (2 mentions) 8w10e array, by Applied Biophysics (1 mentions) Ritc dextran, by Merck Group (1 mentions) Atral, by Merck Group (1 mentions)

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ECIS) system (56 citations) ECIS Zθ (25 citations) Zθ system (4 citations) ECIS Zθ (Theta) system controller (3 citations)

17 protocols using ecis zθ system

1

BEC Resistance Assay for Wnt Signaling

Cited 2 times
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Primary mouse BECs were purified from wild-type and Apcdd1−/− brains as described (Daneman et al., 2010a (link)), plated on poly-D-lysine- and Collagen IV-coated gold electroarray 96-well plates (Applied Biophysics), and grown in endothelial cell media supplemented with growth factors and 10% FBS media (Cell Biologics) to maximum resistance. Cells were switched to 1% FBS and the resistance recorded every 30 min for 48 hr using the ECIS Z-θ system (Applied Biophysics). In some experiments either Wnt3a (250 ng/mL) or 5 μM XAV-939, a tankyrase inhibitor that blocks Wnt signaling activation (Distler et al., 2013 (link); Lim et al., 2017 (link)), were added to a subset of wild-type or Apcdd1−/− BECs when cells were switched to media with low 1% FBS, and the resistance recorded every 30 min for 48 hr using the ECIS Z-θ system (Applied Biophysics). Resistance curves were generated using GraphPad Prism software. Areas under the curve were quantified for the final 48 hr after cells reached maximum resistance using GraphPad Prism software.
Mazzoni J., Smith J.R., Shahriar S., Cutforth T., Ceja B, & Agalliu D. (2017). The Wnt Inhibitor Apcdd1 Coordinates Vascular Remodeling and Barrier Maturation of Retinal Blood Vessels. Neuron, 96(5), 1055-1069.e6.
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2

Real-time Impedance Monitoring of hTM Cells

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Real-time impedance of hTM monolayers was measured using the electric cell-substrate impedance sensing (ECIS) Zθ system (Applied BioPhysics), as described (Wang et al., 2016 (link)). In brief, cells were seeded on 8W10E+ arrays coated with rat collagen before seeding. Once a stable monolayer was established, the cells were exposed to test agents (TREK-1 activators and inhibitors) or control media, and the resistance was measured in each well for 1 h at 10-s intervals in response to a 4-kHz AC frequency (40 electrodes per well). Resistance for each well was normalized to the baseline resistance before the addition of agonist/antagonist to account for baseline differences in the electrodes. Treatment wells were normalized to the control resistance at each time point to determine change in resistance relative to control.
Yarishkin O., Phuong T.T., Bretz C.A., Olsen K.W., Baumann J.M., Lakk M., Crandall A., Heurteaux C., Hartnett M.E, & Križaj D. (2018). TREK-1 channels regulate pressure sensitivity and calcium signaling in trabecular meshwork cells. The Journal of General Physiology, 150(12), 1660-1675.
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3

Quantification of Monolayer Resistance via ECIS

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The resistance of monolayers of 1610 cells was quantified using an ECIS Zθ system (Applied Biophysics, Troy, NY) over a range of 62.5 to 4000 Hz, previously demonstrated to reflect intercellular junctional resistance (Moy et al., 2000 (link); Tiruppathi et al., 1992 (link)). Briefly, 1610 cells (400 µl/well, 2–10 × 104 cells/ml) were plated on 8W10E + 8-well dishes (Applied Biophysics, Troy, NY) where each well was equipped with 40 gold electrodes, and their impedance measured every 90 s for 3 hr. Resistance was quantified as the real component of the impedance and intercellular junctional resistance calculated as the average resistance between 62.5 and 4000 Hz.
Veeraraghavan R., Hoeker G.S., Alvarez-Laviada A., Hoagland D., Wan X., King D.R., Sanchez-Alonso J., Chen C., Jourdan J., Isom L.L., Deschenes I., Smyth J.W., Gorelik J., Poelzing S, & Gourdie R.G. (2018). The adhesion function of the sodium channel beta subunit (β1) contributes to cardiac action potential propagation. eLife, 7, e37610.
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4

Evaluating Resveratrol's Effects on LPS-Induced Epithelial Barrier Dysfunction

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MLE-15 cells (5×104 per well) were seeded in 8-well array plates (Applied BioPhysics, New York, NY) and allowed to grow at 37°C with 5% CO2 in an incubator until confluence. When the cells reached confluence, 20ng/ml LPS-containing medium was added having 50µM RES (LPS+res) or VEH (LPS+veh) and resistance values (Ohm.cm-1) were recorded using ECISzθ system (Applied BioPhysics, New York, NY) every 5 min up to 48 h. Multi-current frequencies were also recorded by using multi-frequency test mode (MFT). Then the acquired data were analyzed to evaluate the effects of RES on the barrier function of LPS-injured epithelial cells (40 (link)).
Alharris E., Mohammed A., Alghetaa H., Zhou J., Nagarkatti M, & Nagarkatti P. (2022). The Ability of Resveratrol to Attenuate Ovalbumin-Mediated Allergic Asthma Is Associated With Changes in Microbiota Involving the Gut-Lung Axis, Enhanced Barrier Function and Decreased Inflammation in the Lungs. Frontiers in Immunology, 13, 805770.
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5

Trans-endothelial Barrier Regulation by PLGF and VEGFR1

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HRECs and HRP were seeded and co-cultured at a 2:1 ratio on the 8-well cultureware (PC, 8W10E). Trans-endothelial electrical resistance (TEER) changes were measured in real-time with the electrical cell-impedance sensing system (ECIS)-Zθ system (Applied Biophysics, NY). The ECIS software-embedded mathematical model of impedance change was used to calculate the TEER (Ω/cm2), measuring the cell-cell barrier and cell-matrix resistance 40 (link). The single-frequency model (4000 Hz) measured resistance and impedance with a 300s interval. After the resistance stabilized and reached a platform, indicating the formation of confluent monolayer and functional barrier, The various treatments: IgG control, PlGF antibody (50 μg/ml), and PlGF antibody (50 μg/ml) + VEGFR1 antibody (50 μg/ml) was added to the medium and then continued to culture for two days. The normalized resistance values were expressed as a percentage relative to vehicle control.
Asakawa J.I., Kodaira M., Nakamura N., Satoh C, & Fujita M. (1999). Chimerism in humans after intragenic recombination at the haptoglobin locus during early embryogenesis. Proceedings of the National Academy of Sciences of the United States of America, 96(18), 10314-10319.
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6

Endothelial Cell Electrical Impedance Assay

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For ECFC-Control and ECFC-CD45, 40,000 cells/well were seeded on fibronectin-coated (0.1mg/cm2) 96 well array with 10 interdigitated electrodes per well (96W10idfPET ECIS array, Applied BioPhysics, NY) in Endothelial Cell Growth Media-2. Once cells adhered and formed a monolayer (~3 hour), media was switched to reduced serum media (2% FBS) and monitored for ~20 hours. For mitral VEC, 10,000 cells/well were seeded on 1% gelatin coated 96W10idfPET ECIS array. When maximum TEER was reached (~23hours), TGFβ1 (1ng/ml) was added, and monitoring continued for ~55 hours. TEER of either ECFCs or mitral VECs was measured at multiple frequencies (62.5–64,000 Hz) in real-time using an ECIS™ Zθ system (Applied BioPhysics). All ECIS measurements are reported at 4,000 Hz, which is the optimal resistance for endothelial cells. The percent change in barrier was quantified compared to appropriate control cells at indicated timepoints. Four technical replicates were included for each condition; data are representative of three independent experiments.
Nasim S., Wylie-Sears J., Gao X., Peng Q., Zhu B., Chen K., Chen H, & Bischoff J. (2023). CD45 is sufficient to initiate EndMT in human endothelial cells. Arteriosclerosis, thrombosis, and vascular biology, 43(5), e124-e131.
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7

Evaluating Endothelial Barrier Disruption by Amyloid-Beta

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Cerebrovascular endothelial barrier formation was assessed using the ECIS Zθ system (Applied Biophysics). All experimental procedures were performed in 8‐well ECIS (8WE10+, Applied Biophysics) gold plated arrays pre‐treated according to the manufacturer's instructions. A monodisperse solution of CMECs was seeded and monitored for 48 h until the electrical resistance reached a plateau at a frequency of 4000 Hz, indicative of barrier formation. At this point, the cell monolayers were treated with 10 µM solutions of the different Aβ peptides or pre‐aggregated Aβ42 in EBM‐2 media containing 1% FBS and followed for 72 h post‐treatment. Barrier permeability was assessed as a decrease in barrier resistance at 4000 Hz compared with untreated cells.
Parodi‐Rullán R., Ghiso J., Cabrera E., Rostagno A, & Fossati S. (2020). Alzheimer’s amyloid β heterogeneous species differentially affect brain endothelial cell viability, blood‐brain barrier integrity, and angiogenesis. Aging Cell, 19(11), e13258.
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8

Monitoring Cell Resistance on Gold Electroarray

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Cells were plated on poly-D-lysine and Fibronectin and Collagen IV-coated gold electroarray 96-well plates (Applied Biophysics) at day 8 of their initial differentiation. Cells were grown in the presence of media and growth factors described above until they reached maximum resistance for 6 d. The resistance was recorded every 3 h for 6 d using the ECIS Z-θ system (Applied Biophysics). Resistance curves were generated using GraphPad Prism software.
Lu T.M., Houghton S., Magdeldin T., Durán J.G., Minotti A.P., Snead A., Sproul A., Nguyen D.H., Xiang J., Fine H.A., Rosenwaks Z., Studer L., Rafii S., Agalliu D., Redmond D, & Lis R. (2021). Pluripotent stem cell-derived epithelium misidentified as brain microvascular endothelium requires ETS factors to acquire vascular fate. Proceedings of the National Academy of Sciences of the United States of America, 118(8), e2016950118.
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9

ECIS Zθ System for Cell Adhesion Measurement

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The high throughput ECIS Zθ system (Applied BioPhysics, Inc.) was used for measurements of adhesion. To measure resistance, well plates were used that contained gold electrode surfaces (Fig. 2B). These surfaces were coated with cysteine to stabilize the electrodes by treatment with 10 mM cysteine for 10 minutes. A 96 well plate with electrodes in the bottom of each well was placed in the well station, and resistance measurements were taken at a frequency of 40,000 Hz. During experiments, resistance, impedance, and capacitance readings were measured. As cells attached to the surface and spread the resistance readings increased.
Spencer A, & Baker A.B. (2016). High Throughput Label Free Measurement of Cancer Cell Adhesion Kinetics Under Hemodynamic Flow. Scientific Reports, 6, 19854.
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10

Quantifying IL-7-Mediated Cell Migration

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Cell migration was detected using an ECIS Zθ system (Applied Biophysics Inc., Troy, NY, USA) as described previously (63 (link)). Briefly, cells were added to 96-well electrode arrays (96W1E) in identical numbers (80,000 cells/well) and allowed to form a fully confluent monolayer. Following confluence, the cells were wounded through the application of 6 V for 30 sec/well, generating a consistently sized ‘wound’ in the monolayer. The change in resistance was measured in each well as the cells migrated back to recolonise the electrode. This process was completed in the presence of varying concentrations of rhIL-7 (0, 1, 10 and 100 ng/ml) and the rate of change of resistance was taken as an indication of cellular migration. Subsequently, 20 ng/ml of rhIL-7 was used in conjunction with a range of small molecule inhibitors to determine potential interactions between these signalling pathways on IL-7-regulated migration.
BARTLETT A., SANDERS A.J., RUGE F., HARDING K.G, & JIANG W.G. (2016). Potential implications of interleukin-7 in chronic wound healing. Experimental and Therapeutic Medicine, 12(1), 33-40.
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