E plate

Manufactured by Agilent Technologies

Sourced in United States, Germany, China

The E-Plate is a laboratory instrument designed for real-time cell analysis. It provides a platform for continuous monitoring and measurement of cellular responses in vitro. The E-Plate utilizes electrical impedance technology to track changes in cell number, morphology, and adhesion in a non-invasive manner.

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Close spelling variants of this product

16-well E-plates (22 citations) E-plates 16 (14 citations) RTCA E-plates (9 citations) Specialized 96-well E-plates (5 citations) E-plates 96 (4 citations) E-Plates L8 (4 citations) E-16-well plates (4 citations) E-plates L16 (3 citations) 8-well E-plates (3 citations) ACEA e-plates (2 citations)

130 protocols using e plate

Real-Time Assessment of NK Cell Cytotoxicity

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The cytotoxicity of NK cells was assessed with a real-time cell analysis (RTCA) assay. Adherent target MCF-7 (3 × 10⁴ cells/well), MDA-MB-231 (3 × 10⁴ cells/well), or SK-BR-3 (4 × 10⁴ cells/well) cells were seeded onto 16-well E-Plates (ACEA Biosciences) in 150 μL of standard RPMI-1640 full medium. For nonadherent K562 cells, 16-well E-Plates were precoated with the Liquid Tumor Killing Assay (anti-CD71) Tethering Kit (ACEA Biosciences) according to the manufacturer's recommendations. K562 cells were seeded onto 16-well E-Plates at a cell density of 1.5 × 10⁴ cells/well. The proliferation of target cells was monitored in the incubator at 37°C (5% CO₂, 95% humidity) for 24 hours with the xCELLigence impedance-based RTCA system (Acea Biosciences). The next day, 100 μL of the medium was aspirated and replaced with the medium containing effector cells (human primary NK cells or NK-92 cell lines) at different effector to target (E:T) ratios. For antibody-dependent cell cytotoxicity (ADCC) assays, trastuzumab (anti-HER2 antibody) was added to appropriate wells at a final concentration of 10 μg/mL. The cells were monitored for the next 20 to 24 hours. Analysis was performed using RTCA Software Pro (ACEA Biosciences). The impedance changes (cell index) were normalized to the end value of the target cells' proliferation and plotted over time as normalized cell index.

Klopotowska M., Bajor M., Graczyk-Jarzynka A., Kraft A., Pilch Z., Zhylko A., Firczuk M., Baranowska I., Lazniewski M., Plewczynski D., Goral A., Soroczynska K., Domagala J., Marhelava K., Slusarczyk A., Retecki K., Ramji K., Krawczyk M., Temples M.N., Sharma B., Lachota M., Netskar H., Malmberg K.J., Zagozdzon R, & Winiarska M. (2021). PRDX-1 Supports the Survival and Antitumor Activity of Primary and CAR-Modified NK Cells under Oxidative Stress. Cancer Immunology Research, 10(2), 228-244.

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Real-Time Monitoring of Cancer Cell Adhesion

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Growth medium derived from sCEA-stimulated and non-stimulated HNNFb cultures were collected and used to monitor the effect of secreted factors on the adhesion and proliferation of cancer cells in real time using an xCELLigence RTCA DP label-free, impedance-based cell-sensing instrument (ACEA Biosciences Inc.; San Diego, CA). Briefly, HeLa and CEA-expressing HeLa.CEA cells were suspended in culture medium and dispensed (2.0 × 10⁴ cells per well) into 16-well sensor plates (E-plates; ACEA Biosciences Inc.) pre-equilibrated with growth medium (serum-free or supplemented with FBS (10%; v/v)) or conditioned medium from stimulated and non-stimulated HNNFb cultures to assess the effects of HNNFb stimulation by sCEA on the adhesion of CEA-expressing cancer cells. Changes in adherence and proliferation of CEA null and CEA-expressing colon cancer cells were also determined by dispensing LS174T, HT-29, MC38 cells (2.0 × 10⁴ cells per well) into sensor plates (E-plates; ACEA Biosciences Inc.) pre-equilibrated with conditioned medium from stimulated and non-stimulated HNNFb cultures. Changes in relative impedance were measured at 1-min intervals, over the course of 48 hours [2 (link)–5 (link), 20 (link)].

Abdul-Wahid A., Cydzik M., Fischer N.W., Prodeus A., Shively J.E., Martel A., Alminawi S., Ghorab Z., Berinstein N.L, & Gariépy J. (2018). Serum-derived carcinoembryonic antigen (CEA) activates fibroblasts to induce a local re-modeling of the extracellular matrix that favors the engraftment of CEA-expressing tumor cells. International journal of cancer, 143(8), 1963-1977.

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Cytotoxicity Assessment of Solvents

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In brief, 2,000 cells were added into wells of E-plates (ACEA Biosciences, San Diego, CA, USA). After 24 hours of culture, cells were treated with DMSO, methanol and ethanol at the concentrations of 10%, 5%, 2.5%, 1.25%, 0.6%, 0.3%, 0.15%, and 0%. The E-plates were then connected into the xCelligence RTCA machine (ACEA Biosciences) in the incubator for the next 48 hours. Cell proliferation was measured by recording the change in the impedance of electron flow caused by adherent cells. The experiments were done in triplicate (n=3). The data were analyzed by the software attached to the xCelligence RTCA system (ACEA Biosciences).

Pham P.V., Nguyen H.T.L., & Truong K.D. (2020). Comparative cytotoxic effects of methanol, ethanol and DMSO on human cancer cell lines. Biomedical Research and Therapy, 7(7), 3855-3859.

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Real-Time Cell Proliferation Assay

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In this assay, 2×10⁴ cells were plated in E-plate (ACEA Biosciences, Inc., a division of Agilent, USA) with 150 μL DMEM containing 10% FBS. The E-plate was placed in an RTCA instrument (ACEA Biosciences, Inc., a division of Agilent, USA) in a standard CO₂ cell culture incubator for 100 or 60 h.

Li D., Ni X.F., Tang H., Zhang J., Zheng C., Lin J., Wang C., Sun L, & Chen B. (2020). KRT17 Functions as a Tumor Promoter and Regulates Proliferation, Migration and Invasion in Pancreatic Cancer via mTOR/S6k1 Pathway. Cancer Management and Research, 12, 2087-2095.

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Real-time Eosinophil Impedance Monitoring

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Real-time monitoring of eosinophil detachment and cell death were assessed by measuring electrical impedance using the xCELLigence Real-Time Cell Analyzer (ACEA Biosciences, San Diego CA) as previously described (26 (link), 27 (link)). Briefly, media was placed in 96-well gold electrode coated plates (E-plates, ACEA Biosciences), allowed to equilibrate and a background reading was obtained. Eosinophils were then seeded at 1 × 10⁶ cells/100 μl and allowed to settle for ~5 hours. SP-A was added at various concentrations and changes in electrical impedance were measured over time. Impedance measurements, presented as a normalized “Cell Index,” are calculated as detailed (26 (link), 27 (link)). Under these conditions, a loss of Cell Index is associated with eosinophil detachment and cytotoxicity. Individual traces of cytotoxicity over time were averages of 2 – 3 technical replicates; standard deviations were eliminated for clarity. Quantification of cytoxicity was accomplished by measuring the area under the curve (AUC) after normalization of cell index. Graphical AUC measurements include baseline correction for untreated cells to best display changes.

Dy A.B., Arif M.Z., Addison K.J., Que L.G., Boitano S., Kraft M, & Ledford J.G. (2019). Genetic Variation in Surfactant Protein A2 Delays Resolution of Eosinophilia in Asthma. Journal of immunology (Baltimore, Md. : 1950), 203(5), 1122-1130.

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In vitro Extravasation Assay for Tumor Cell Invasion

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In vitro extravasation assay was performed as previously described [18 ]. Briefly, 2.5 × 10⁴ HUVEC cells in 100 µl were seeded in E-plates (ACEA xCelligence). Once HUVEC monolayer formed (19–21 h), 1 × 10⁴ MDA-MB-231 cells were added on top of the monolayer. A decrease in cell index indicates invasion through the HUVEC monolayer by tumor cells. Penetration of HUVEC monolayer was monitored for up to 10 h by using xCelligence Real-time Cell Analyzer (Acea Biosciences).

Siddiqui A., Gollavilli P.N., Schwab A., Vazakidou M.E., Ersan P.G., Ramakrishnan M., Pluim D., Coggins S., Saatci O., Annaratone L., HM Schellens J., Kim B., Asangani I.A., Rasheed S.A., Marchiò C., Sahin O, & Ceppi P. (2019). Thymidylate synthase maintains the de-differentiated state of triple negative breast cancers. Cell Death and Differentiation, 26(11), 2223-2236.

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Real-Time T Cell Killing Assay

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To assess real time T cell killing of mesothelioma cell line (H28, H2452) we used the iCELLigence RTCA instrument (ACEA Biosciences). First, 50 μl of cell culturing media was added to each well of 16 well E-Plates (ACEA Biosciences) to measure the background impedance. Next, target cells were seeded at a density of 10,000 (H28) or 20,000 (H2452) cells/well of the E-Plate in a volume of 50 μl. The following day, when the Cell Index (CI) reached a level of around 1, effector cells (cultured CD8+ T cells) were added at an effector to target (E:T) ratio of 1:10 in 100 μl (reaching a total of 200 μl/well). Data recordings are shown as Normalized Cell Index.

Chiaro J., Antignani G., Feola S., Feodoroff M., Martins B., Cojoc H., Russo S., Fusciello M., Hamdan F., Ferrari V., Ciampi D., Ilonen I., Räsänen J., Mäyränpää M., Partanen J., Koskela S., Honkanen J., Halonen J., Kuryk L., Rescigno M., Grönholm M., Branca R.M., Lehtiö J, & Cerullo V. (2023). Development of mesothelioma-specific oncolytic immunotherapy enabled by immunopeptidomics of murine and human mesothelioma tumors. Nature Communications, 14, 7056.

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Real-time monitoring of cell proliferation

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Cells were seeded at 5000 cells per well in 200 μl of complete media in E-plates (ACEA Biosciences, San Diego, CA, USA) and grown overnight while monitored with an xCELLigence DP system (ACEA Biosciences) which monitors cellular events in real time by measuring electrical impedance across interdigitated gold micro-electrodes integrated on the bottom of tissue culture plates [16 (link), 17 ]. Cells were washed three times with PBS and cultured with 180 μl EGM-2 basal media (no growth factors or supplements) and incubated for a minimum of 6 h before further treatment. Treatments were prepared at 10 × concentrations and added to each well in a total volume of 20 μl. The xCELLigence DP recorded cell index readings every 15 min for 3 days after treatment. Cell index readings were normalized before treatment and cell proliferation ratios were determined from four biological replicates and represent the relative numbers of cells compared to control cells. A two-way ANOVA with Holm–Sidak’s multiple comparisons test was used to compare IPSE treatment to medium-alone control, with P ≤ 0.05 deemed significant.

Mbanefo E.C., Agbo C.T., Zhao Y., Lamanna O.K., Thai K.H., Karinshak S.E., Khan M.A., Fu C.L., Odegaard J.I., Saltikova I.V., Smout M.J., Pennington L.F., Nicolls M.R., Jardetzky T.S., Loukas A., Brindley P.J., Falcone F.H, & Hsieh M.H. (2020). IPSE, an abundant egg-secreted protein of the carcinogenic helminth Schistosoma haematobium, promotes proliferation of bladder cancer cells and angiogenesis. Infectious Agents and Cancer, 15, 63.

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Real-time Cell Proliferation Assay

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The rate of cell proliferation was assessed in real-time using an xCELLigence RTCA dual-plate instrument (ACEA Biosciences, San Diego, CA, USA) as previously described [21 (link)]. Briefly, HaCaT (8000 cells/well) and SiHa (6000 cells/well) were seeded into 100 µL of DMEM with 10% FBS in E-plates (ACEA Biosciences). The impedance value, expressed as cell index, was recorded every 30 min. Cells were treated with 50 µL of the indicated concentration of crinamine or 0.5% DMSO in growth medium when they reached log phase and impedance was recorded for another 96 h. The cell proliferation rate was derived from the slope of the line between two given time points.

Khumkhrong P., Piboonprai K., Chaichompoo W., Pimtong W., Khongkow M., Namdee K., Jantimaporn A., Japrung D., Asawapirom U., Suksamrarn A, & Iempridee T. (2019). Crinamine Induces Apoptosis and Inhibits Proliferation, Migration, and Angiogenesis in Cervical Cancer SiHa Cells. Biomolecules, 9(9), 494.

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Real-time Cell Confluency Monitoring

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Cells were plated to equal confluence on E-plates in triplicate (ACEA biosciences Inc.). At 24 h after plating, cells were treated with indicated drug doses and analysed on the xCELLigence RTCA MP system with impedance readings taken every 30 min to 4 h. Cells were retreated with drugs every 48–72 h and impedance measurements were taken to calculate the relative quantity of confluence for attached cells to the bottom of each well.

Towers C.G., Guarnieri A.L., Micalizzi D.S., Harrell J.C., Gillen A.E., Kim J., Wang C.A., Oliphant M.U., Drasin D.J., Guney M.A., Kabos P., Sartorius C.A., Tan A.C., Perou C.M., Espinosa J.M, & Ford H.L. (2015). The Six1 oncoprotein downregulates p53 via concomitant regulation of RPL26 and microRNA-27a-3p. Nature Communications, 6, 10077.

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