The 8W10E arrays (IBIDI, Munich, Germany) were treated with 200 µL 10 mM L-cysteine dissolved in sterile water for 15 min at room temperature. Wells were washed 3× with 300 µL sterile water. Then, 200 µL PureCol Type 1 bovine collagen solution (Advanced Biomatrix, San Diego, CA, USA) diluted 1 in 100 in sterile water, was added to wells and incubated at 37°C overnight. Wells were washed 3× with BEGM. Then, 400 µL BEGM was added to each well and the array was stabilised in the electric cell-substrate impedance sensing (ECIS) Zθ instrument (Applied Biophysics, Troy, NY, USA). Medium was replaced with 7.5×104 cells in BEGM. The array was run at multiple frequencies (including the specific frequencies of 400 Hz and 32 kHz) for 72 h continuously on the ECIS Zθ instrument, as described previously [14 (link)].
8w10e arrays
Manufactured by Ibidi
Sourced in Germany
The 8W10E arrays are a type of lab equipment designed for electrochemical analysis. They provide a platform for conducting parallel electrochemical measurements on up to 8 individual working electrodes. The core function of the 8W10E arrays is to enable efficient and high-throughput electrochemical experimentation and data collection.
Automatically generated - may contain errors
Lab products found in correlation
Ecis zθ instrument, by Applied Biophysics (2 mentions)
Embryomax, by Merck Group (2 mentions)
Purecol type 1 bovine collagen solution, by Advanced BioMatrix (1 mentions)
Ecis 1600r, by Applied Biophysics (1 mentions)
Accutase, by Merck Group (1 mentions)
Ecis zθ theta instrument, by Applied Biophysics (1 mentions)
4 protocols using 8w10e arrays
1
ECIS-based Analysis of Cell Adhesion on Collagen
2
Assessing Cell Monolayer Integrity
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Cells (1.5 × 105) grown in 8W10E arrays (Ibidi) were monitored using an ECIS1600R (ECIS, electric cell–substrate impedance sensing) instrument (Applied BioPhysics, Inc) (48 (link)), and ECIS software, version 1.2.186.0 PC. An increase in capacitance at 64 kHz mirrors a loss of monolayer integrity translating as cell detachment and death.
3
Endothelial Barrier Function Assessment via ECIS
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Endothelial barrier function was assessed by Electric Cell-substrate Impedance Sensing (ECIS) using the ECIS Zθ instrument (Applied BioPhysics, ibidi). iECs were grown to confluence on 0.1% gelatine (EmbryoMax, Sigma Aldrich) coated 8W10E arrays (ibidi) (3 × 10 4 cells/ well) for 72 h. Cells were starved for 5 h before stimulation with VEGFA (5 nM) and PBS as control. Barrier function was assessed by ECIS measurement of resistance at 4000 Hz.
4
Quantifying Endothelial Cell Adhesion and Spreading
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Cell adhesion and spreading was quantified by recording capacitance at a frequency of 64 kHz, where the decrease in capacitance is directly proportional to the electrode coverage, using the ECIS Zθ (theta) instrument (Applied BioPhysics) [127 ]. ECs were harvested with Accutase (Sigma Aldrich) and pelleted by 4 min of 300 rpm centrifugation. Pellets were resuspended in basal M199 media. The amount of 5 × 104 cells were seeded on 0.1% gelatine (EmbryoMax, Sigma Aldrich) coated 8W10E arrays (ibidi, Germany). To interfere with β1-integrin–ECM interactions, ECs were subjected to β1-integrin–blocking antibody (2.5 μg/ml, Abcam, P5D2 #ab24693) to isotype control IgG1 (2.5 μg/ml, Cell Signaling, #5415). Cells were kept in suspension and incubated with antibodies at 37°C by gentle inversion of the tube every 5 min. After 30 min, cells were seeded as described earlier. Influence of SMKI on cell adhesion and spreading was analyzed by culturing ECs for 3 d in M199 basal growth medium in the presence of K02288 (1 μM), SB-431542 (10 μM), or DMSO as control without medium change. Cells were harvested and seeded on arrays as described earlier.
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