Impedance measurement (xCELLigence® technology)
This biosensor technology uses electrical impedance to allow continuous real-time monitoring of cellular activity in a non-invasive label-free manner. The impedance can be frequently measured (30 seconds being the minimum interval between two measurements), which allows an accurate characterisation of both early and late events of the cellular response.
Cellular Impedance Technical Overview (xCELLigence technology)
The xCELLigence® system uses custom designed plates, which have a high-density gold electrode array upon which the target cells adhere and grow. The xCELLigence® instruments are placed in a standard CO2 cell culture incubator and interfaces via a cable with analysis and control units located outside the incubator. Cells adhere to the plate surface and modify the electrical impedance across the array, which is measured and recorded by the xCELLigence® software. The impedance values are converted by the software into the Cell Index (CI). Any response that induces changes in cell morphology (size, volume, shape or spreading), cell number (proliferation or death) or movement (migration or invasion) can be investigated using xCELLigence® technology. User real-time cell analysis (RTCA) software allows for real-time control and monitoring of the instrument, and includes real-time data display and analysis functions.
Site Baclesse
On the Baclesse site, the ImpedanCELL platform is located in the research building of the CCC with a dedicated 30 m2 air conditioned cell culture lab that was recently opened to provide an optimal service for users. The cell culture room is equipped with with two microbiological safety station, three CO2 incubators (170 L for the reception of the two xCELLigence® RTCA MP stations and 230 L for one IncuCyte S3) and all the small equipment required (multichannel pipettes, centrifuge, etc.). The platform shares the premises with the BioTICLA laboratory of the Inserm UMR 1086 ANTICIPE (10 m2 storage space, 30 m2 for preparation, 150 m2 lab area, 50 m2 office space and a 14 m2 meeting room). In addition, external users can use an office (14 m2) equipped with a workstation and software dedicated to the analysis of results.
xCELLigence® RTCA MP system
The ImpedanCELL platform is equipped with two xCELLigence® RTCA MP (Multiple-Plate) systems that can host up to six-well electronic microtiter plates each allowing the high-throughput study of 1200 conditions simultaneously with a measure every 30 seconds on several days. This MP system allow to obtain various information on the cellular activity (adhesion, proliferation, viability, morphology).
Each RTCA MP system is composed of 4 items:
Cells placed in the upper chamber of the apparatus pass through the microporous membrane (Boyden chamber) and the impedance signal is measured on the bottom part of the membrane.
The ImpedanCELL platform is equipped with an xCELLigence® DP (Dual-Purpose) system to perform chemotactic migration/invasion assays. This instrument also allow to study the cellular activity (adhesion, proliferation, viability, morphology), however, with a low throughput.
The RTCA DP system is composed of 3 items:
In parallel, the platform offer an access to an automated, ultrafast multiplex and non-invasive brightfield and fluorescence imaging with multiparameter image analysis (CELLAVISTA® RS HighEnd, SYNENTEC GmbH) using dedicated E-Plate VIEW 96 (each well contains an electrode-free viewing strip to facilitate microscopy). A daily monitoring of cellular morphology offer complementary information, which gives clues about the nature of the observed phenomenon (adhesion, cellular detachment, proliferation, cell death, epithelial-mesenchymal transition, differentiation, etc.). CELLAVISTA® RS HighEnd is equipped with 5 different lenses (x2, x4, x10, x20 and x40) and 6 fluorescent (FL) excitation channels and 6 detection wavelengths which covers approx. 80% of all FL dyes. (Hoechst/DAPI, eGFP, mCherry and Alexa®647).
These equipment were acquired thanks to the support of the CCC F. Baclesse, European funding (ERDF) and the University of Caen Normandy.
Site LABÉO
The Equine Health research platform (Hippolia) is equipped with a third xCELLigence® RTCA MP (Multiple-Plate) available on the LABÉO site in a BSL-2 confinement room, allowing to perform complementary experiments in the field of virology and bacteriology. This instrument can can host up to six-well electronic microtiter plates each allowing the high-throughput study of 600 conditions simultaneously with a measure every 30 seconds on several days. This MP system allow to obtain various information on the cellular activity (adhesion, proliferation, viability, morphology) and to follow in real-time the cytopathic effect after virus infection.
Each RTCA MP system is composed of 4 items:
These equipment were acquired thanks to the support of LABÉO of the combined State-Region funding scheme
for the equine sector CENTAURE and European funding (ERDF).
Live cell imaging (IncuCyte® S3)
In 2017, the ImpedanCELL platform acquired two live cell imaging systems (IncuCyte® S3), one for each site. This equipment automates image acquisition and analysis in long-term kinetics (phase contrast and fluorescence), directly in a CO2 incubator, and allows the study of six 96 well plates (or other formats such as 25 or 75 cm2 culture flasks) simultaneously. It is equipped with an automated optical turret with three objectives (x4, x10, x20) and allows visualization in phase contrast and in fluorescence (green: Ex- 440-480 nm, Em - 504-544 nm and red: Ex - 565-605 nm, Em - 625-705 nm) to monitor several types of experiments in parallel. These two systems were acquired in order to increase the versatility and the ability of both sites to respond to requests for investigations of high-throughput real-time cellular activity (action of molecules on spheroids, wound healing, characterization of cell death, observation of cytopathic effect after virus infection, etc.) using add-on software modules (Chemotaxis, NeuroTrack, Scratch Wound/Migration, Angiogenesis, Spheroid) and/or Live-Cell Imaging reagents (Caspase-3/7 and Annexin V Apoptosis Assays, Nuclei labelling…) developed by the manufacturer or by the ImpedanCELL platform.
These equipment were acquired thanks to the support of the combined State-Region funding scheme INNOVONS (Baclesse site), the combined State-Region funding scheme for the equine sector CENTAURE (LABÉO site) and the University of Caen Normandy.
