Automated Cell Migration

ECIS® instruments include an elevated field mode allowing for electroporation and wounding. The ECIS® wound is precisely defined, as it includes only those cells on the electrode. Additionally, with ECIS® the ECM protein coating is not scraped off and is unaffected by the current. Example publication; Peitzman et al., (2015) "Agonist Binding to β-Adrenergic Receptors on Human Airway Epithelial Cells Inhibits Migration and Wound Repair." American Journal of Physiology - Cell Physiology 309(12):C847 doi:10.1152/ajpcell.00159.2015

Barrier Function

Epithelial cells and endothelial cells regulate the passage of molecules across cell layers. Diseases, especially vascular disease, occur when this function is impaired. ECIS® provides a highly sensitive real-time continous TEER measurement ideal for these types of studies. Example publication; Kim et al., (2015) "Critical Role of Sphingosine-1-Phosphate Receptor-2 in the Disruption of Cerebrovascular Integrity in Experimental Stroke." Nature Communications 6:7893 doi:10.1038/ncomms8893

Signal Transduction

ECIS® is especially useful to monitor the signal transduction pathways activated by G protein cou-pled receptors (GPCR). GPCR activation, regardless of the second messenger, results in alterations of the cell’s cytoskeletal elements, causing morpholog-ical changes. Example publication; Shinde et al., (2013) "STIM1 Controls Endothelial Barrier Function Independ-ently of Orai1 and Ca2+ Entry." Science Signaling 6(267):ra18. doi:10.1126/scisignal.2003425

Cell Proliferation

As cells proliferate two factors act to change the impedance: cell number and cell morphology. In most instances the cells grow asynchronously and the impedance gradually increases until a maximum when cells become confluent. The impedance change is approximately linear with cell number while the cells are sub-confluent. Example publication; MacKenzie et al., (2013) "Triptolide Induces the Expres-sion of miR-142-3p: A Negative Regulator of Heat Shock Protein 70 and Pancreatic Cancer Cell Proliferation." Molecular Cancer Therapeutics 12(7):1266 doi:10.1158/1535-7163.MCT-12-1231

Differentiation and Stem Cell Biology

When cells differentiate they change their behavior allowing ECIS® to follow the events of cell differen-tiation. While most tools available to characterize stem cells preclude their further use, the label-free non-invasive nature of ECIS® allows for subsequent use of characterized stem cells. Example publication; Alvarez et al., (2011) "The Hedgehog Pathway Promotes Blood-Brain Barrier In-tegrity and CNS Immune Quiescence." Science (New York, N.Y.) 1727(6063):1727 doi:10.1126/science.1206936

Cell Toxicity

The ECIS® system has been used specifically to assess the cytotoxicity of a variety of toxicants. ECIS-based toxicity tests are far superior to simple cell death assays, because cell function is also monitored. Example publication; Banerjee et al., (2013) "Triptolide-Induced Cell Death in Pancreatic Cancer Is Medi-ated by O-GlcNAc Modification of Transcription Factor Sp1." The Journal of Biological Chemistry 288(47):33927 doi:10.1074/jbc.M113.500983

Cell Attachment and Spreading

Traditional "counting attached cells assays" can only quantify the number of cells attached to any ECM coating. ECIS® assays give feedback on the strength of the attachment of the cells to the ECM. Example publication; Kourtidis et al., (2015) "Distinct E-Cadherin-Based Complexes Regulate Cell Behaviour through miRNA Processing or Src and p120 Catenin Activity." Nature Cell Biology 17(9):1145 doi:10.1038/ncb3227

Cell Invasion

ECIS® can distinguish between transmigration mechanisms that leave the monolayer intact from those that disrupt the cell layer. Published examples include metastatic cell and leukocyte trans-endothe-lial migration, as well as the migration of pathogens such as yeast, anthrax, streptococcus, plasmodium, trypanosomes, and spirochetes. Example publication; Goc et al., (2013) "P21 Activated Kinase-1 (Pak1) Promotes Prostate Tumor Growth and Microinvasion via Inhibition of Transforming Growth Factor Expression and Enhanced Matrix Metalloproteinase 9 Secretion." The Journal of Biological Chemistry 288(5):3025 doi: 10.1074/jbc.M112.424770

Inflammation

ECIS recovery-after-wounding assays allow for the discovery of molecules which aid in the process of tissue repair. ECIS barrier function assays specifically measure the response of epithelial and endothelial cells to secreted cytokines and can give indirect information about the binding of immune cells to the epithelium or endothelium. Example publication; Migliorini et al., (2013) "The Antiviral Cytokines IFN-α and IFN-β Modulate Parietal Epithelial Cells and Promote Podocyte Loss: Implications for IFN Toxicity, Viral Glomerulonephritis, and Glomerular Regeneration." The American Journal of Pathology 183(2):431. doi:10.1016/j.ajpath.2013.04.017

Cancer Metastasis

The study of cancer metastasis incorporates many of the applications above in ways that reflect the specific biology of cancer. "Electric Cell-Substrate Impedance Sensing and Cancer Metastasis" is volume 17 of the book series "Cancer Metastasis - Biology and Treatment" edited by Wen G. Jiang and contains 12 chapters entirely devoted to the use of ECIS® in cancer metastasis. With an introductory chapter by Ivar Giaever it is an excellent resource for using ECIS® in cancer research. The book "Electric Cell-Substrate Impedance Sensing and Cancer Metastasis" is available from Springer Netherlands

Design Your Own Assay by Quantifying Cell Behavior

Cell function modulates cell morphology. ECIS® is capable of detecting and quantifying morphology changes in the sub-nanometer to micrometer range. In ECIS® a small alternating current (I) is applied across the electrode pattern at the bottom of the ECIS® arrays (direct current cannot be used). This results in a potential (V) across the electrodes which is measured by the ECIS® instrument. The impedance (Z) is determined by Ohm’s law Z = V/I.

When cells are added to the ECIS® Arrays and attach to the electrodes, they act as insulators increasing the impedance. As cells grow and cover the electrodes, the current is impeded in a manner related to the number of cells covering the electrode, the morphology of the cells and the nature of the cell attachment. When cells are stimulated to change their function, the accompanying changes in cell morphology alter the impedance. The data generated is impedance versus time.

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