Rtca sp instrument

Manufactured by Roche

Sourced in Switzerland, Germany

The RTCA SP instrument is a real-time cell analysis system that measures impedance changes in cell cultures over time. It provides a label-free, non-invasive method to monitor cell proliferation, cytotoxicity, and cell-substrate interactions in real-time.

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Lab products found in correlation

Xcelligence real time cell analyzer system, by Roche (1 mentions) Rtca software, by Roche (1 mentions) E plates 16, by Agilent Technologies (1 mentions) Rtca software version 1, by Roche (1 mentions) E plates 96, by Agilent Technologies (1 mentions)

6 protocols using rtca sp instrument

Real-Time Monitoring of Cell Behavior Using xCELLigence RTCA

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The xCELLigence real-time cell analyzer (RTCA) system (Roche and ACEA Biosciences) is a label-free and non-invasive assay system based on a continuous measurement of electrical impedance to monitor biological processes of living cells in real time. The impedance is affected by the cells attached on the electrodes and gives information about their biological status, including cell number, viability, morphology and the degree of cell adhesion [16 (link)].
Growth of the HBEC lines was continuously monitored for at least 24 h using 96-well plates that contain microelectrodes (E-plate 96) and the RTCA SP instrument (Roche, Rotkreuz, Switzerland). The measurement of the impedance background was performed with 100 µl of cell culture medium per well. After adding the cell suspension the final volume in a single well was 200 μL (10,000 cells/well). Electric impedance was recorded every 30 min.
The impedance is displayed as Cell Index (CI), a relative and dimensionless value directly influenced by cell attachment, spreading and cell proliferation.

Stempin S., Engel A., Winkler N., Buhrke T, & Lampen A. (2015). Morphological and molecular characterization of the human breast epithelial cell line M13SV1 and its tumorigenic derivatives M13SV1-R2-2 and M13SV1-R2-N1. Cancer Cell International, 15, 110.

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Measuring Cell Impedance Dynamics

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Cell function (hMEC) was assessed by changing of electrical impedance measured by the xCELLigence system, using the RTCA SP instrument (Roche Applied Science) as previously described^{42 (link)}. Briefly, cells at a density of 2 × 10⁴ per well were plated in a 96 well E-plate containing gold microelectrode sensors on the bottom to measure cellular impedance inside each well. Cells were allowed to adhere overnight and after were treated with rS1p for 60 min. Values are represented by cell index numbers, which are a dimensionless unit of measurement representing the measurement of zero impedance when cells are absent and increasing as cells adhere, spread and divide, and analysed using RTCA software (Roche). The slope was measured based on the normalised cell index from the point in which rS1p treatment was administered.

Montezano A.C., Camargo L.L., Mary S., Neves K.B., Rios F.J., Stein R., Lopes R.A., Beattie W., Thomson J., Herder V., Szemiel A.M., McFarlane S., Palmarini M, & Touyz R.M. (2023). SARS-CoV-2 spike protein induces endothelial inflammation via ACE2 independently of viral replication. Scientific Reports, 13, 14086.

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Real-Time Monitoring of MDA-MB-231 Cell Growth

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For monitoring the growth of MDA-MB-231 cells in real-time (RTCA), a realtime cell analyzer (xCELLigence RTCA SP Instrument, ROCHE) was employed. After the background of E-plate reading (100 μL medium), the cells were added to E-plate (3 × 10³ cells/well; 50 μL) and grown for 22−24 h. Then, the investigated compounds at various concentrations were added to 50 μL of media. The impedance was monitored for additional 4 days. An arbitrary unit CI (cell index) is a quantitative measure reflecting the status of the cells (number of attached cells and cell status such as morphology) in an electrode-containing well. Normalized CI at a certain time point is calculated as CI at the time point divided by CI at a reference time point.

Kasparkova J., Kostrhunova H., Novohradsky V., Logvinov А.A., Temnov V.V., Borisova N.E., Podrugina T.A., Markova L., Starha P., Nazarov A.A, & Brabec V. (2022). Novel cis-Pt(II) Complexes with Alkylpyrazole Ligands: Synthesis, Characterization, and Unusual Mode of Anticancer Action. Bioinorganic Chemistry and Applications, 2022, 1717200.

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Real-Time Cell Cytotoxicity Assay

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All experiments were performed using the respective target cell culture medium. A 50-μl aliquot of the medium was added to E-Plates 16 (ACEA Biosciences, San Diego, CA) for measurement of background values. Target cells were seeded in the medium at a density of 10,000 cells/well. Suitable cell densities were determined by prior titration experiments. Cell attachment was monitored using an RTCA SP instrument (Roche) with RTCA software, version 1.1 (Roche), until the plateau phase was reached, which usually required ∼22–26 hours T cells were added at different effector to target (E:T) ratios, ranging from 20:1 to 5:1. Upon addition of effector cells, impedance measurements were performed every 15 minutes for up to 81 hours. All experiments were performed in duplicate. Changes in electrical impedance were expressed as a dimensionless cell index (CI) value, which was derived from relative impedance changes corresponding to cellular coverage of the electrode sensors, normalized to baseline impedance values with medium only. To analyze the acquired data, CI values were exported and the percentage of lysis was calculated in relation to control cells lacking any effector T cells. Percent lysis was determined using the following formula at various time points: (CI tumor only−[CI tumor+T cells])/(CI tumor only)×100.16

Matsui H., Hazama S., Tamada K., Udaka K., Irie A., Nishimura Y., Miyakawa T., Doi S., Nakajima M., Kanekiyo S., Tokumitsu Y., Shindo Y., Tomochika S., Yoshida S., Iida M., Suzuki N., Takeda S., Yamamoto S., Yoshino S., Ueno T, & Nagano H. (2019). Identification of a Promiscuous Epitope Peptide Derived From HSP70. Journal of Immunotherapy (Hagerstown, Md. : 1997), 42(7), 244-250.

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Measuring Cell Proliferation Dynamics

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The xCELLigence system RTCA SP instrument (Roche Applied Science) monitors changes in the cell index (a measure of cell attachment to the plate) which has been shown to effectively correlate to proliferation, adhesion, and viability changes⁶⁴. To assess proliferation cells were seeded in a 96-well gold electrode sensor plate (E-plates 16) and monitored every 15 min for varying times (50–100 h) in 10% FBS. Cell index (CI) values were normalized 7 h after plating (to exclude the adhesion phase). Cell growth was calculated as the slope (hours-1) of the cell index curve during a total of 50 h. In no case was cell death due to excessive confluence as confirmed by plate inspection with a microscope.

Cabezudo S., Sanz-Flores M., Caballero A., Tasset I., Rebollo E., Diaz A., Aragay A.M., Cuervo A.M., Mayor F J.r, & Ribas C. (2021). Gαq activation modulates autophagy by promoting mTORC1 signaling. Nature Communications, 12, 4540.

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RPTEC/TERT1 Cell Impedance Monitoring

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If not otherwise indicated, RPTEC/TERT1 cells were seeded on E-plates 96 (ACEA Biosciences Inc., San Diego, CA, USA) at 30% density and placed into the RTCA SP instrument (Roche Diagnostics, Mannheim, Germany). After a growth and maturation period of at least 14 days, cells were treated with compounds or solvent control (0.5% DMSO) or left non-treated. Cellular impedance was measured in intervals of 5 min for a total of 14 days. Treatment was renewed on Mondays, Wednesdays, and Fridays. Cell indices obtained for treated cells were normalized to non-treated cells and are given as percent.

Secker P.F., Schlichenmaier N., Beilmann M., Deschl U, & Dietrich D.R. (2019). Functional transepithelial transport measurements to detect nephrotoxicity in vitro using the RPTEC/TERT1 cell line. Archives of toxicology, 93(7).

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