As cells adhere to the biocompatible gold microelectrodes that line the well bottoms in an E-Plate they impede the flow of electric current between electrodes. Using our proprietary algorithm, this impedance signal is converted to a specific parameter called Cell Index. The Cell Index is an excellent measure of what the cells are actually doing over time: proliferating, spreading, changing shape, dying, responding to specific stimuli, etc. The Cell Index measurement has been reviewed and accepted in over 700 peer-reviewed publications.
No. The electric field is extremely weak and non-invasive. Repeated studies at ACEA Biosciences and elsewhere have confirmed the technique to be harmless to the cells.
For all seven xCELLigence models the Stations that are placed inside the incubator are specifically designed to withstand the high temperature and humidity of laboratory incubators.
The range is approximately 30 meters in an open environment.
If the Control Unit is out of the WiFi range of the iCELLigence Station the data will not be lost and the experiment will not be interrupted. Once an experiment is started, the Station records and saves all the data. The saved data automatically synchronizes to the Control Unit once the WiFi connection is re-established. Data collected prior to a power outage will not be lost as long as the outage is less than 15 minutes in length.
Yes, each well is measured individually, in sequential fashion. Since the RTCA Instrument measures essentially the entire bottom surface of the well, the dynamic range of the system approaches 2 logs of cell growth – from 100 cells per well to confluence (depending on cell type). Also, well-to-well precision and accuracy are excellent. In our laboratory we typically achieve well-to-well CVs of less than 10%.
A: Yes. Cells can be nicely imaged directly on the E-Plate L8, E-Plate View 16, and E-Plate View 96 devices through the electrode-free viewing area.
Yes. E-Plates can be coated with any number of matrices to enhance or prevent cell attachment. Examples of amenable surface coatings include poly-L-lysine, fibronectin, and collagen IV.
E-Plates are made of biocompatible materials and are tissue-culture treated at the time of manufacture. They are sterile, and designed for single use. Cell growth on E-Plates is essentially identical to what is observed on most standard cell culture plates.
Like conventional cell culture plates, the E-Plate is not designed or intended for reuse.
Cells attach to the planar gold electrode sensor array at the bottom of the wells. These electrodes cover approximately 70-80% of the well bottom surface area. All components of the E-Plates are biocompatible, sterile, and tissue-culture treated.
RTCA Applications and Data
The time-dependent curves generated by the RTCA Instrument yield a wealth of information about the actual kinetics of cellular events occurring in the wells. Cell morphology changes yield curves which differ distinctly from those resulting from changes in cell number. The use of appropriate control treatments and/or orthogonal assays during initial xCELLigence assay development make it easy to differentiate between cell morphology change and cell proliferation in an RTCA curve. Typically, however, simply viewing the cells in a microscope is sufficient for making this distinction.
The RTCA systems are used for a broad range of research applications, including cell proliferation, cytotoxicity, cell adhesion, RTK, GPCR, RNAi functional analyses, natural killer cell activity, ADCC, CDC, virus neutralizing antibody detection, and bacterial toxin neutralizing antibody detection. This list is not exhaustive. For a more comprehensive list of applications, please click the “Applications” tab within each of the seven xCELLigence product pages.
Hundreds of cell lines as well as some primary cells have been tested. Most adherent cell types can be analyzed on the RTCA System. Cells which are not naturally adherent have also been successfully studied by tethering them to the well bottom using antibodies. In the absence of this special treatment, non-adherent cells generally cannot be detected by the sensors and thus cannot be measured very well directly. However, in certain experiments this can be a benefit. This is especially true for cell-mediated cytotoxicity, where the killing of adherent target cells by suspension effector cells (Natural Killer, Cytotoxic T-lymphocytes, etc.) is being measured. For an exhaustive list of cells that have been successfully analyzed using xCELLigence, please download our list of verified cell lines.
RTCA has been used so extensively over the past 10 years that a massive body of data (over 700 publications) and experience now enable the correlation of RTCA curve features with distinct biological processes (such as cytoskeletal rearrangement upon GPCR stimulation or the induction of apoptosis by a DNA damaging agent). Importantly, if different drugs or treatments induce the same biological phenomena they give rise to similar RTCA curves. For this reason, a novel drug’s mechanism of action (MOA) can often be inferred simply by comparing the RTCA curve it induces with a panel of RTCA curves from drugs whose MOA is known. Most importantly, appropriate controls and/or orthogonal assays can be used to link the features of an RTCA curve with specific cellular/biochemical processes.
Videos: Product Overviews
Videos: Research Presentations
Dr. Felix Bohne
Institute of Virology, German Research Center for Environmental Health, Helmholtz Zentrum München
Dr. Sarah K. Lamore
AstraZeneca Pharmaceuticals, Waltham, MA
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Dr. Keith L. Knutson
Mayo Clinic, Jacksonville, FL
The Vaccine & Gene Therapy Institute of Florida, Port Saint Lucie, FL
Dr. Melissa L. Fishel
Indiana University School of Medicine, Indianapolis, IN
Dr. Matthew F. Peters
AstraZeneca Pharmaceuticals, Boston, MA