Epithelial volt ohm meter evom2

Manufactured by World Precision Instruments

Sourced in United States, United Kingdom

The Epithelial Volt/Ohm Meter EVOM2 is a laboratory instrument designed to measure the transepithelial electrical resistance (TEER) and voltage across epithelial cell monolayers. It provides precise measurements of these parameters, which are important indicators of the integrity and permeability of the epithelial barrier.

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Lab products found in correlation

Stx2 chopstick electrode, by World Precision Instruments (3 mentions) Transwell insert, by Corning (2 mentions) Enspire multilabel plate reader, by PerkinElmer (1 mentions) Caco 2 cells, by Sumitomo Pharma (1 mentions) Htb 37, by American Type Culture Collection (1 mentions) Perfluorocarbon, by Thermo Fisher Scientific (1 mentions) Texas red dextran, by Thermo Fisher Scientific (1 mentions) Synergymx multi mode microplate reader, by Agilent Technologies (1 mentions) Lactate assay kit, by Merck Group (1 mentions) Fv10i w, by Olympus (1 mentions) Fluoview software, by Olympus (1 mentions) Stx2 chopstick electrode set, by World Precision Instruments (1 mentions)

29 protocols using epithelial volt ohm meter evom2

Measuring Epithelial Cell Integrity

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The integrity of the cell monolayer was routinely tested by measuring TEER with the Epithelial Voltohmmeter EVOM 2 (World Precision Instruments, Sarasota, FL, USA) according to manufacturer’s protocol and as previously discussed [18 (link)].

Piccapane F., Gerbino A., Carmosino M., Milano S., Arduini A., Debellis L., Svelto M., Caroppo R, & Procino G. (2021). Aquaporin-1 Facilitates Transmesothelial Water Permeability: In Vitro and Ex Vivo Evidence and Possible Implications in Peritoneal Dialysis. International Journal of Molecular Sciences, 22(22), 12535.

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Transepithelial Electrical Resistance Measurements

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For TER measurements, cells were plated at confluency in Transwell inserts and grown in NC medium. TER was measured in confluent cell monolayers subjected to a Ca²⁺ switch. In this case, rapid TJ disassembly, as assessed by obtaining background TER values, was induced by incubating confluent cell monolayers for 1 h in Ca²⁺-deprived LC medium containing 1 mM EGTA; cells were then switched back to NC medium for the indicated time, as described previously (Nunbhakdi-Craig et al., 2002 (link)). In other experiments, cells were plated at confluency in Transwell inserts and cultured for 72 h in NC medium to allow complete TJ maturation. Transfected protein expression was induced with sodium butyrate as soon as cells were attached. For each time point, TER was measured using an Epithelial Volt/Ohm Meter (EVOM²; World Precision Instruments). Particular care was taken to measure TER under strictly similar experimental conditions across cell lines (Sheller et al., 2017 (link)). TER values (Ω.cm²) were normalized to the filter area of the monolayer and calculated by subtracting blank values from the filter and bathing medium (Nunbhakdi-Craig et al., 2002 (link)).

Schuhmacher D., Sontag J.M, & Sontag E. (2022). A Novel Role of PP2A Methylation in the Regulation of Tight Junction Assembly and Integrity. Frontiers in Cell and Developmental Biology, 10, 911279.

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Measuring Epithelial Barrier Integrity

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The cells in transwell culture were daily monitored by measuring the TEER using the Epithelial Voltohmmeter (EVOM2, World Precision Instruments, Inc., Sarasota, FL). The chopstick electrodes were placed in both the apical and basal compartments with the tip fully immersed in the media for the measurement. Each transwell was measured thrice at three different positions, and the mean value was taken and then multiplied with the total surface area of the transwell. For each condition, an average of three replicate transwell readings was used to plot the TEER graph. The principles of TEER measurement are comprehensively illustrated in a review by Benson et al.^{16 (link)}

Ghatak S., Reveiller M., Toia L., Ivanov A.I., Zhou Z., Redmond E.M., Godfrey T.E, & Peters J.H. (2015). Bile Salts at Low pH Cause Dilation of Intercellular Spaces in In Vitro Stratified Primary Esophageal Cells, Possibly by Modulating Wnt Signaling. Journal of gastrointestinal surgery : official journal of the Society for Surgery of the Alimentary Tract, 20(3), 500-509.

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Transepithelial Electrical Resistance Measurement

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According to the manufacturer’s instructions, the TEER was evaluated with the Epithelial Volt/Ohm Meter EVOM2 (World Precision Instruments, Hertfordshire). Electrodes were equilibrated in media and placed in the upper and lower chambers. Background measurement of a Matrigel-coated insert (with no cells) was subtracted from the reading and the value was multiplied by the growth surface area, to determine the TEER values (Ohms*cm²).

Clé M., Constant O., Barthelemy J., Desmetz C., Martin M.F., Lapeyre L., Cadar D., Savini G., Teodori L., Monaco F., Schmidt-Chanasit J., Saiz J.C., Gonzales G., Lecollinet S., Beck C., Gosselet F., Van de Perre P., Foulongne V., Salinas S, & Simonin Y. (2021). Differential neurovirulence of Usutu virus lineages in mice and neuronal cells. Journal of Neuroinflammation, 18, 11.

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Measuring Epithelial Barrier Function

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A confluent monolayer of SMG-C6 cells was grown in a 24-well Corning Transwell™ filter (6.5-mm diameter, 0.4-μm pore size). Transepithelial electrical resistance (TER) was measured using an Epithelial Volt Ohm Meter EVOM2 (World Precision Instruments, Sarasota, FL, USA). All TER values were obtained from at least three wells. Final values were calculated by subtracting the blank filter value (80 Ω) and multiplying by the surface area of the filter. For the paracellular tracer flux assay, 1 mg/mL 4-kDa (68059, Sigma-Aldrich, St. Louis, MO, USA) or 40-kDa FITC-dextran (53379, Sigma-Aldrich) was added to the medium in the basal compartment, the medium in the apical compartment was collected after an incubation at 37 °C for 3 h, and the paracellular tracer flux was measured using EnSpire Multilabel Plate Reader (PerkinElmer, Waltham, MA, USA).

Huang Y., Liu H.M., Mao Q.Y., Cong X., Zhang Y., Lee S.W., Park K., Wu L.L., Xiang R.L, & Yu G.Y. (2021). High Glucose Reduces the Paracellular Permeability of the Submandibular Gland Epithelium via the MiR-22-3p/Sp1/Claudin Pathway. Cells, 10(11), 3230.

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Caco-2 cell monolayer transepithelial transport

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Caco-2 cells (DS Pharma Biomedical Co., Ltd., Tokyo, Japan) were maintained in high-glucose Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum, 100 U/mL penicillin G potassium, 100 μg/mL streptomycin, and 1% nonessential amino acids [50 (link)]. Cells were plated on a 24-well Transwell plate or 6-well Transwell plate at a density of 8 × 10⁴ cells/cm² and incubated in a CO₂ incubator at 37 °C for 21 days. Experiments were performed using cells that had previously been passaged 50 to 55 times. The integrity of the monolayer was measured by determining the TEER with an Epithelial Voltohmmeter (EVOM2, World Precision Instruments, Sarasota, FL, USA) [51 (link)].

Ikarashi N., Nagoya C., Kon R., Kitaoka S., Kajiwara S., Saito M., Kawabata A., Ochiai W, & Sugiyama K. (2019). Changes in the Expression of Aquaporin-3 in the Gastrointestinal Tract Affect Drug Absorption. International Journal of Molecular Sciences, 20(7), 1559.

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Caco-2 Cell Monolayer Transepithelial Resistance

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UBM sheets from each group were cut to 8 mm diameter circles, placed into transwell inserts (Corning^™ BioCoat^™ Assay System, 1.0um membrane) with BMC luminal surface facing upwards and sterilized in situ with ETO gas (16 h cycle at 50 °C in a Series 3plus EOGas Sterilizer, Anderson Sterilizers, Inc., Haw River, NC). For transepithelial electrical resistance measurements Caco-2 cells (passages 24–28, ATCC^® HTB-37^™) were cultured to approximately 80% confluence in MEM containing non-essential amino acids, 1mM sodium pyruvate, and 20% FBS. Caco-2 cells were seeded on the BMC luminal surface and the functional response was evaluated using a rapid differentiation system (Corning^™ Biocoat^™ HTS Caco-2 Assay) per manufacturer’s instructions. The functional response was measured by transepithelial electrical resistance (TEER). TEER of Caco-2 monolayers was measured with an Epithelial Voltohmmeter (EVOM2, World Precision Instruments). Electrical resistance of each scaffold type without cells was also measured and subtracted from the total electrical resistance determined with the monolayer to calculate the TEER of the monolayer.

White L.J., Taylor A.J., Faulk D.M., Keane T.J., Saldin L.T., Reing J.E., Swinehart I.T., Turner N.J., Ratner B.D, & Badylak S.F. (2016). The impact of detergents on the tissue decellularization process: a ToF-SIMS study. Acta biomaterialia, 50, 207-219.

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Differentiating iPSCs into RPE Cells

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The iPSC were grown to confluence and the E8 media was changed to bFGF-depleted ES media to initiate spontaneous differentiation. Pigmented foci were manually dissected, dissociated and seeded as previously described^{29 (link)}. For TER measurements, iPSC-derived RPE was seeded on Matrigel-coated BD Falcon cell culture inserts (Dominique Dutscher, Brumath, France). TER was measured using the Epithelial Volt/Ohm Meter EVOM2 (World Precision Instruments, Hertfordshire, U.K.) as described^{29 (link)}.

Torriano S., Erkilic N., Baux D., Cereso N., De Luca V., Meunier I., Moosajee M., Roux A.F., Hamel C.P, & Kalatzis V. (2018). The effect of PTC124 on choroideremia fibroblasts and iPSC-derived RPE raises considerations for therapy. Scientific Reports, 8, 8234.

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Transepithelial Electrical Resistance Measurement

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At P3, the iPSC-derived RPE was cultured on Matrigel-coated, clear BD Falcon cell culture inserts with high density 0.4 μM pores (Dominique Dutscher) in 24-well plates. The TER was measured using the Epithelial Volt/Ohm Meter EVOM2 (World Precision Instruments, Hertfordshire, U.K.) according to the manufacturer's instructions. Briefly, electrodes were sterilized in 70% ethanol for 5 min, rinsed and equilibrated in media, then placed in the compartmentalised chambers with the longer electrode vertically touching the bottom of the dish in the lower chamber and the shorter electrode in the upper chamber without touching the cell layer. TER was recorded once the value stabilised, approximately 5 s after placing the electrode. To calculate the final TER values (Ohms·cm²), the background measurement of a Matrigel-coated insert without cells was subtracted from the reading and the value multiplied by the growth surface area.

Simonin Y., Erkilic N., Damodar K., Clé M., Desmetz C., Bolloré K., Taleb M., Torriano S., Barthelemy J., Dubois G., Lajoix A.D., Foulongne V., Tuaillon E., Van de Perre P., Kalatzis V, & Salinas S. (2018). Zika virus induces strong inflammatory responses and impairs homeostasis and function of the human retinal pigment epithelium. EBioMedicine, 39, 315-331.

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Airway Epithelial Electrophysiology Assay

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HBECs were exposed to pooled 15 μL BALF per culture for 6 h vs. PBS (control). After this time, perfluorocarbon (Thermo Fisher Scientific, Waltham, MA, USA, cat no. 394890500) was added to prevent evaporation and membrane potentials were recorded using a KCl-filled glass microelectrode that was positioned in the ASL with a micromanipulator and a KCl/agar macroelectrode that was positioned in the serosal media. Transepithelial voltages were then recorded using an FD223 electrometer (World Precision Instruments, Sarasota, FL, USA) as described [23 (link),26 (link)]. Changes in voltages caused by CFTR-mediated transepithelial Cl⁻ secretion and ENaC-mediated transepithelial Na absorption were determined by calculating bumetanide and amiloride-inhibitable voltages, respectively. We then measured resistance using an Epithelial Volt Ohm Meter (EVOM2, World Precision Instruments, Sarasota, FL, USA) as described [48 (link)]. Equivalent open circuit currents (I_eqs) were then calculated according to Ohm’s law.

Ghosh A., Coakley R.D., Alexis N.E, & Tarran R. (2023). Vaping-Induced Proteolysis Causes Airway Surface Dehydration. International Journal of Molecular Sciences, 24(20), 15348.

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