TER was measured with an EVOM Volt-Ohm meter (EVOL, World Precision Instruments, Aston, Herts, UK) with STX2 electrode (World Precision Instruments, Inc., Sarasota, FL, USA). Briefly, cells were added into the 0.4-μm pore size insert (Greiner Bio-One Ltd., Stonehouse, UK) and allowed to reach full confluency. Electrodes were placed at the upper and lower chambers, then resistance was measured with the Volt-Ohm meter. Fluorescein isothiocyanate (FITC)-dextran 10 kDa and TRITC dextran 40 kDa were applied to cells apically. Basolateral dextran passage was analyzed with a GloMax®-Multi Microplate Multimode reader (Promega UK Ltd., Southampton, UK).
0.4 μm pore size insert
Manufactured by Greiner
Sourced in United Kingdom
The 0.4-μm pore size insert is a laboratory equipment component designed for filtration purposes. It features a pore size of 0.4 micrometers, which allows for the separation and isolation of particles or molecules of specific sizes. This product is intended to be used as a part of a larger laboratory setup or system, without further interpretation of its intended use.
Automatically generated - may contain errors
Lab products found in correlation
Evom voltohmmeter, by World Precision Instruments (4 mentions)
Glomax multi microplate multimode reader, by Promega (4 mentions)
Black 96 well cell culture microplate, by Greiner (3 mentions)
Stx2 chopstick electrode, by World Precision Instruments (3 mentions)
Stx2 electrode, by World Precision Instruments (1 mentions)
4 protocols using 0.4 μm pore size insert
1
Transepithelial Electrical Resistance and Dextran Permeability
2
Transepithelial Resistance and Permeability Assay
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An EVOM Voltohmmeter (World Precision Instruments), equipped with STX2 chopstick electrodes (World Precision Instruments) was used to measure the transepithelial resistance (TER). Briefly, 5 × 104 cells were plated into a 0.4-μm pore size insert (Greiner Bio-One Ltd) and cultured to 100% confluence. Electrodes were placed in the upper and lower chambers and resistance was subsequently measured using a Volt-Ohm meter. Inserts without cells in medium were set as a blank control. Following the analysis of TER, the medium in the upper chambers was replaced with normal medium containing 0.2 mg/ml fluorescein isothiocyanate (FITC)-dextran 10 kDa. Then, 50 µl medium from outside the insert was transferred into a black 96-well cell culture microplate (Greiner Bio-One) in duplicate every 2 h for 10 h. The basolateral dextran passage was analysed using a GloMax®-Multi Microplate Multimode reader (Promega Corporation), with an excitation wavelength of 490 nm and an emission wavelength of 510–570 nm. Each measurement was normalized to the 0 h via subtraction.
3
Measuring Epithelial Barrier Integrity
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TER was used to assess the integrity of tight junction dynamics in cell culture models of epithelial monolayers as a widely accepted quantitative technique. An EVOM voltohmmeter (World Precision Instruments, Aston, Herts, UK), equipped with STX2 chopstick electrodes (World Precision Instruments, Inc., Sarasota, FL, USA) was used to measure the TER. Briefly, 5×10 4 cells were seeded into a 0.4 μm pore size insert (Greiner Bio-One Ltd, Stonehouse, UK) and allowed to reach full confluence. Following the replacement with fresh medium, electrodes were placed at the upper and lower chambers and resistance were measured with the Volt-Ohm meter. Inserts containing cell-free medium were set as blank control. After TER was recorded, the medium in the upper chamber was replaced with normal medium containing 0.2 mg/ml fluorescein isothiocyanate (FITC)-dextran 10 kDa. 50 μl of medium from outside of the insert was transferred into a black 96-well cell culture microplate (Greiner Bio-One) in duplicate every 2 hours for 10 hours. Basolateral dextran passage was analysed with a GloMax®-Multi Microplate Multimode Reader (Promega UK Ltd., Southampton, UK) at excitation 490nm and emission 510-570nm. Measurement of dextran-indicated PCP was then normalized to the 0 h via subtraction.
4
Quantifying Epithelial Barrier Integrity
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TER was used to assess the integrity of tight junction dynamics in cell culture models of epithelial monolayers as a widely accepted quantitative technique. An EVOM voltohmmeter (World Precision Instruments, Aston, Herts, UK), equipped with STX2 chopstick electrodes (World Precision Instruments, Inc., Sarasota, FL, USA) was used to measure the TER. Briefly, 5×10 4 cells were seeded into a 0.4 μm pore size insert (Greiner Bio-One Ltd, Stonehouse, UK) and allowed to reach full confluence. Following the replacement with fresh medium, electrodes were placed at the upper and lower chambers and resistance were measured with the Volt-Ohm meter. Inserts containing cell-free medium were set as blank control. After TER was recorded, the medium in upper chamber was replaced with normal medium containing 0.2 mg/ml fluorescein isothiocyanate (FITC)-dextran 10 kDa. 50 μl of medium from outside of the insert was transferred into a black 96-well cell culture microplate (Greiner Bio-One) in duplicate every 2 hours for 10 hours. Basolateral dextran passage was analysed with a GloMax®-Multi Microplate Multimode Reader (Promega UK Ltd., Southampton, UK) at excitation 490nm and emission 510-570nm. Measurement of dextran-indicated PCP was then normalized to the 0 h via subtraction.
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