NCI-H441 cells were seeded onto a 12-mm Transwell with a 0.4 μm pore polyester membrane insert (Corning CoStar Corp., Cambridge, MA, USA) at a density of 2.0 × 104 cells/cm2. TEER was measured using EVOMX (World Precision Instruments, Sarasota, FL, UK) and expressed as Ohm*cm2 after subtracting the resistance of a blank Transwell insert.
Evomx
Manufactured by World Precision Instruments
Sourced in United States, Germany
The EVOMX is a digital electrochemical workstation designed for a wide range of electrochemical measurements. It features high-performance potentiostat and galvanostat capabilities for techniques such as voltammetry, amperometry, and electrochemical impedance spectroscopy.
Automatically generated - may contain errors
Lab products found in correlation
18 gauge sharp needle, by Terumo (2 mentions)
Epithelial voltohmmeter, by World Precision Instruments (1 mentions)
Endohm chamber, by World Precision Instruments (1 mentions)
Stx2 electrode, by World Precision Instruments (1 mentions)
Ketalar, by Daiichi Sankyo (1 mentions)
Celactal, by Bayer (1 mentions)
9 protocols using evomx
1
Measuring Epithelial Cell Barrier Integrity
2
Evaluating Epithelial Barrier Function
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To evaluate epithelial barrier function, trans-epithelial electrical resistance (n = 10) was measured using an Epithelial Volt-ohm meter and Ag/AgCl electrodes (EVOMX, World Precision Instruments, Sarasota, FL). The trans-epithelial electrical resistance of each sample was measured before (TER1) and after (TER2) the epithelia were removed by exposure to 0.02 M EDTA for 1 hour. Finally, the TER of each sample was calculated as follows: TER = TER1-TER2.
3
Corneal Transepithelial Electrical Resistance Measurement
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Corneal TER was measured as previously described [16] (link). Briefly, a 1.0-mm-diameter Ag/AgCl electrode (Physiotech, Tokyo, Japan) was inserted into the anterior chamber through a small incision in the peripheral cornea, which had been made with an 18-gauge sharp needle (Terumo, Tokyo, Japan). Using biomedical adhesive (Alon-Alpha A; Sankyo), a 6.0-mm-internal diameter (0.28-cm2 inner area) nitrile rubber O-ring (Union Packing; SAN-EI, Osaka, Japan) was fixed on the cornea. Then, 60 μL of HBSS was placed inside the ring at the center of the cornea, and the other electrode was carefully placed on the cornea. The TER was measured using a volt-ohm meter (EVOMX, World precision Instruments, Saeasota, FL), which generates ±20-μA alternating current (AC) square wave current at 12.5 Hz, and data were recorded using a thermal arraycorder (WR 300-8; Graphtech, Tokyo, Japan).
4
Measuring Epithelial Cell Integrity
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TEER measurements were done prior to medium change using an epithelial tissue voltohmeter (EVOMX; World Precision Instruments) fitted with chopstick electrodes (STX-2) and surface-normalized values calculated according to the manufacturer’s instructions. Measurements across inserts with medium but no cells served as control, and resulting values were subtracted from results with cells.
5
Measuring Endothelial Barrier Integrity
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TEER measurements were performed as previously described [27 (link)]. The electrical resistance across the endothelial monolayer was assessed using an EndOhm™ chamber coupled to a resistance meter (EVOMX; World Precision Instruments, Inc, Sarasota, FL, USA). The TEER readings were collected before (0 h) and after (2 and 4 h) extracts addition. After subtracting the values of empty inserts (without cells and extracts) and multiplying by the area of the insert (1.12 cm2), TEER values expressed as Ω × cm2 were obtained and the percentage of variation from average control readings (without extracts) were determined. After the last reading, the media from both compartments were collected and used for the transport studies mentioned above, and then the empty inserts were used for the following permeability assay.
6
Transepithelial Electrical Resistance of hiPSC-RPE
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For functional assessment, TER of hiPSC-RPE monolayers cultured on gfr-Matrigel-coated transwell filters was measured using an epithelial voltohmmeter (EVOMX) following the manufacturer’s instructions (World Precision Instruments, Berlin, Germany). Briefly, electrodes were sterilized with 70 % ethanol, rinsed in 150 mM NaCl solution and placed in the transwell filter with the longer voltage electrode positioned in the lower chamber touching the bottom of the dish while the shorter current electrode was placed in the upper chamber. Net TER was calculated from TER recordings by subtracting background measurements obtained from gfr-Matrigel-coated transwell filters without cells. TER measurements were obtained by multiplying net TER values with the surface area of the transwell filters and reported as Ω*cm2.
7
Evaluating Metformin's Impact on Airway Epithelial Barrier
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After receiving SmallAir™ 3D airway epithelia comprising fully differentiated human primary cells and following the vendor’s protocol (49 ), they were transferred to 24-well plates pre-filled with 700 μL of SmallAir™ media in the basal side to create the ALI. After 3 days of incubation, 1000 μM solution of SD metformin (25% PR) was added to the cells. TEER values were measured using EVOMX (Epithelial VoltOhmMeter) and electrode (STX2) (World Precision Instruments, Sarasota, FL). To measure TEER, 200 μL of the cell media were added to the apical surface of the inserts. The long part of the electrode was inserted through the gap of the insert and leaned on the bottom of the well, and the short stem was above in the apical surface, inside the culture media. TEER values were obtained before exposure to the drug solution and after exposure to them. The response was measured after 3 h of exposure and then every 24 h for 5 days. Every time, the TEER measurement was finished, the media was removed from the apical surface in order to leave the cells in ALI conditions.
8
Measuring Transepithelial Electrical Resistance of hiPSC-RPEs
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9
In Vivo Corneal Transepithelial Resistance Measurement
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Rabbits were anesthetized using an intramuscular injection of 30 mg/kg ketamine (Ketalar, Daiichi Sankyo, Tokyo, Japan) and 5 mg/kg xylazine (Celactal, Bayer Yakuhin, Osaka, Japan). A 1.0 mm diameter custom-made Ag/AgCl electrode (Physiotech, Tokyo, Japan) was inserted into the anterior chamber, following a small incision made in the peripheral cornea using an 18-gauge sharp needle (Terumo, Tokyo, Japan). A 6.0 mm internal diameter (0.28 cm 2 inner area) nitrile rubber O-ring (Union Packing, SAN-EI, Osaka, Japan) was fixed on the cornea using biomedical adhesive (Alon-Alpha A, Daiichi Sankyo, Tokyo, Japan). Subsequently, 80 µL of HBSS were added in the ring, with a second electrode then placed in the HBSS on the cornea. The TER was measured in real time using a volt-ohm meter (EVOMX, World Precision Instruments, Sarasota, FL, USA) generating a ±20 μA AC square wave current at 12.5
Hz. The specific methodology and photographs of the in vivo corneal TER measurement system have been previously published [9] [10] [11] [12] (link). The sample size for the corneal TER study was set at 4 to 8, which we found to be sufficient for our statistical analyses in our previous TER studies [9] [10] [11] [12] (link).
Hz. The specific methodology and photographs of the in vivo corneal TER measurement system have been previously published [9] [10] [11] [12] (link). The sample size for the corneal TER study was set at 4 to 8, which we found to be sufficient for our statistical analyses in our previous TER studies [9] [10] [11] [12] (link).
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