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Vcc mc8

Manufactured by Physiologic Instruments
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The VCC MC8 is a multichannel data acquisition system designed for recording and analyzing physiological signals. It provides eight independent analog input channels, allowing for simultaneous data collection from multiple sources. The device features high-resolution analog-to-digital conversion and programmable gain and offset controls to ensure accurate signal capture. The VCC MC8 is suitable for a variety of applications where reliable and precise data acquisition is required.

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Lab products found in correlation

Ussing chamber, by Physiologic Instruments (5 mentions) Dvc 1000, by World Precision Instruments (5 mentions) Acquire and analyze software, by Physiologic Instruments (3 mentions) P2300, by Physiologic Instruments (2 mentions) Powerlab, by ADInstruments (2 mentions) Acquire and analysis 2, by Physiologic Instruments (2 mentions) Forskolin, by Thermo Fisher Scientific (2 mentions) Em csys 8, by Physiologic Instruments (2 mentions) Acquire analyse 2, by Physiologic Instruments (2 mentions) Cftr inhibitor 172, by Merck Group (1 mentions) Acquire and analyze 2, by Physiologic Instruments (1 mentions) Easymount ussing chamber system, by Physiologic Instruments (1 mentions) Labchart, by ADInstruments (1 mentions) Ussing chamber, by Thermo Fisher Scientific (1 mentions) Acquire analyze software, by Physiologic Instruments (1 mentions) Epithelial voltohmmeter, by World Precision Instruments (1 mentions) O dianisidine, by Merck Group (1 mentions) Fitc 4000 fd 4, by Merck Group (1 mentions) Snapwell inserts, by Corning (1 mentions)

22 protocols using vcc mc8

1

Measuring CFTR-mediated Anion Excretion

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Costar Transwell filters (6.5 mm) were used for culturing of F508del-CFTR CF cells and transduction with SFPQ/GFP adenoviral vector or control GFP vectors was performed on the basolateral side of the cell monolayer at 37 °C for 48 h. These filters were then mounted and equilibrated in Physiologic Instruments Ussing chambers as previously described78 (link), with some modifications. In the subsequent step, 10 µM amiloride was added to the apical chamber to inhibit epithelial sodium channel (ENaC)-mediated sodium absorption. After 2 min, 10 µM forskolin was added to both the basolateral and apical chambers to activate CFTR-mediated anion excretion. Following another 2 min, CFTR-mediated anion excretion was inhibited using a CFTR Inhibitor-172 (Sigma-Aldrich) to the apical chamber. During this time, currents had achieved steady-state. Short-circuit current (Isc) and transepithelial resistance (TER) were continuously measured using a Physiologic Instruments VCC-MC8 and Physiologic Instruments Acquire and Analyze 2.3 data acquisition hardware and software. Agonist- or inhibitor-induced changes in short-circuit current (ΔIsc) were calculated from differences in the mean Isc over the 10 s period preceding reagent additions.
Kumar P., Soni D.K., Sen C., Larsen M.B., Mazan-Mamczarz K., Piao Y., De S., Gorospe M., Frizzell R.A, & Biswas R. (2021). SFPQ rescues F508del-CFTR expression and function in cystic fibrosis bronchial epithelial cells. Scientific Reports, 11, 16645.
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2

Ussing Chamber Measurement of Transepithelial Resistance

Cited 1 time
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Inserts were mounted in modified Ussing chambers (P2300; Physiologic Instruments, San Diego, CA) and continuously short circuited with an automatic voltage clamp (VCC MC8; Physiologic Instruments) as described previously (Edinger et al., 2012 (link), 2014 (link)). The apical and basolateral chambers each contained 4 ml of Ringer solution (120 mM NaCl, 25 mM NaHCO3, 3.3 mM KH2PO4, 0.8 mM K2HPO4, 1.2 mM MgCl2, 1.2 mM CaCl2, and 10 mM glucose). Chambers were constantly gassed with a mixture of 95% O2, 5% CO2 at 37°C, which maintained the pH at 7.4 and established a circulating perfusion bath within the Ussing chamber. Simultaneous transepithelial resistance was recorded by applying a 2-mV pulse per minute via an automated pulse generator. Recordings were digitized and analyzed using PowerLab (AD Instruments, Colorado Springs, CO).
LIU X., EDINGER R.S., KLEMENS C.A., PHUA Y.L., BODNAR A.J., LAFRAMBOISE W.A., HO J, & BUTTERWORTH M.B. (2017). A MicroRNA Cluster miR-23–24–27 Is Upregulated by Aldosterone in the Distal Kidney Nephron Where it Alters Sodium Transport. Journal of cellular physiology, 232(6), 1306-1317.
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3

Ussing Chamber Monitoring of MTE Cell Response

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MTE cells on 6.5 mm permeable filters were mounted in an EasyMount Ussing chamber system (Physiologic Instruments, San Diego, CA) and bathed on both sides with Krebs-Ringers Buffer containing: 115 mM NaCl, 25 mM NaHCO3, 0.4 mM KH2PO4, 2.4 mM K2HPO4, 1.2 mM CaCl2, and 1.2 mM MgCl2 with 10 mM glucose. Bath solutions were continuously circulated with a gas lift by bubbling with 95 % air and 5 % CO2 at 37 °C (pH 7.4). MTE cell monolayers were voltage clamped and monitored for changes in short-circuit current (Isc) and TER with a multichannel voltage/current clamp VCC MC8 (Physiologic Instruments, San Diego, CA). MTE cell monolayers were clamped to 0 mV and a 5 mV pulse of 200-msec duration was imposed every 10 s. Changes in Isc (ΔIsc) were calculated from the difference between the initial Isc measurement at baseline and the peak measurement change after adding 2,5-dimethylpyrazine alone or in combination with pharmacological inhibitors/agonists. Data was analyzed using Acquire and Analyze software, version 2.3 (Physiologic Instruments, San Diego, CA).
Sherwood C.L, & Boitano S. (2016). Airway epithelial cell exposure to distinct e-cigarette liquid flavorings reveals toxicity thresholds and activation of CFTR by the chocolate flavoring 2,5-dimethypyrazine. Respiratory Research, 17, 57.
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4

Quantifying CFTR Activity in Differentiated hNECs

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Differentiated hNECs were mounted in circulating Ussing chambers (VCC MC8; Physiologic Instruments, San Diego, CA). Chambers were filled with asymmetric chloride (Cl−) Ringer solution (bicarbonate free) and short-circuit current (Isc, µA/cm2) was measured as previously described (Awatade et al., 2021 (link)). Baseline current was stabilised for 30 min before transepithelial electrical resistance (TEER) measurements were recorded. The following pharmacological compounds were then sequentially added: 100 μM apical amiloride to inhibit epithelial sodium channels, 10 μM apical VX-770 (IVA) or 0.01% DMSO (vehicle) to potentiate CFTR-activated currents, 10 μM basal forskolin to induce cAMP activation of CFTR, 30 μM apical CFTRInh-172 to inhibit CFTR-specific currents and 100 μM apical ATP to activate calcium-activated chloride currents. Using Acquire and Analysis 2.3 software (Physiologic Instruments, San Diego, CA) data recordings of Isc were obtained. Cumulative changes of Isc in response to forskolin and CFTR modulator were used to quantify total CFTR-activated currents. Changes in Isc in response to forskolin alone (no CFTR modulator) were considered baseline activity (ΔIsc-Fsk).
Allan K.M., Astore M.A., Kardia E., Wong S.L., Fawcett L.K., Bell J.L., Visser S., Chen P.C., Griffith R., Jaffe A., Sivam S., Vittorio O., Kuyucak S, & Waters S.A. (2023). Q1291H-CFTR molecular dynamics simulations and ex vivo theratyping in nasal epithelial models and clinical response to elexacaftor/tezacaftor/ivacaftor in a Q1291H/F508del patient. Frontiers in Molecular Biosciences, 10, 1148501.
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5

Measuring Epithelial Ion Transport

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Following the formation of a monolayer, the medium was removed and the cells were rinsed and bathed in buffer solution (in mM) (130 NaCl, 0.47 KCl, 0.124 MgSO4, 0.33 CaCl2, 10 Hepes, 2.5 NaH2PO4, 10 dextrose). Custom made chambers were designed and built to measure short-circuit current in 0.33-cm2 Transwell inserts (SI Appendix, Supplementary Methods). The cells were maintained at 37 °C and short-circuit current was measured using an VCCMC8 multichannel voltage clamp (Physiologic Instruments), and LabChart (ADInstruments) was used to record measurements.
Chang D., Whiteley A.T., Bugda Gwilt K., Lencer W.I., Mekalanos J.J, & Thiagarajah J.R. (2020). Extracellular cyclic dinucleotides induce polarized responses in barrier epithelial cells by adenosine signaling. Proceedings of the National Academy of Sciences of the United States of America, 117(44), 27502-27508.
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6

Differentiated Nasal Epithelium Ussing Chamber Assay

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Snapwell inserts carrying differentiated nasal epithelia were mounted in a vertical diffusion chamber resembling a Ussing chamber with internal fluid circulation. Both apical and basolateral hemichambers were filled with 5 mL of a solution containing (in mM) 126 NaCl, 0.38 KH2PO4, 2.13 K2HPO4, 1 MgSO4, 1 CaCl2, 24 NaHCO3, and 10 glucose. Both sides were continuously bubbled with a gas mixture containing 5% CO2–95% air and the temperature of the solution was kept at 37 °C. The transepithelial voltage was short-circuited with a voltage-clamp (DVC-1000, World Precision Instruments, Sarasota, FL, USA; VCC MC8 Physiologic Instruments, Reno, NV, USA) connected to the apical and basolateral chambers via Ag/AgCl electrodes and agar bridges (1 M KCl in 1% agar). The offset between the voltage electrodes and fluid resistance was adjusted to compensate for the parameters before experiments. The short-circuit current was recorded by analogical to digital conversion on a personal computer.
Sondo E., Cresta F., Pastorino C., Tomati V., Capurro V., Pesce E., Lena M., Iacomino M., Baffico A.M., Coviello D., Bandiera T., Zara F., Galietta L.J., Bocciardi R., Castellani C, & Pedemonte N. (2022). The L467F-F508del Complex Allele Hampers Pharmacological Rescue of Mutant CFTR by Elexacaftor/Tezacaftor/Ivacaftor in Cystic Fibrosis Patients: The Value of the Ex Vivo Nasal Epithelial Model to Address Non-Responders to CFTR-Modulating Drugs. International Journal of Molecular Sciences, 23(6), 3175.
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7

Ussing Chamber Measurements of Epithelial Cells

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Confluent cells grown on Snapwells were mounted in Ussing chambers (Physiologic Instruments, San Diego, California, USA) to measure short circuit current (ISC) and transepithelial electrical resistance (TER). Cells were bathed in Krebs buffer solution (pH 7.4) consisting of 115 mM NaCl, 2 mM KH2PO4, 2.4 mM MgCl2*6H2O, 25 mM NaHCO3, 8 mM KCl, 1.3 mM CaCl2, and supplemented 10 mM glucose and 10 mM mannitol on the basolateral and apical sides, respectively. Experiments with TBCA were performed in Krebs buffer without MgCl2. Tissue was voltage clamped to 0 V and unclamped every 20 seconds and a 5 mV potential difference applied using a voltage-clamp apparatus (VCC MC8, Physiologic Instruments). Change in current was measured using a digital data acquisition system (BioPac, Goleta, CA) and TER calculated using Acquire and Analyze software (Physiologic Instruments). Cell viability was determined at the end of each Ussing chamber experiment through measurement of the change in short circuit current (Isc) induced by 10 μM forskolin (Alfa Aesar, Ward Hill, MA).
Lahey K.A., Ronaghan N.J., Shang J., Dion S.P., Désilets A., Leduc R, & MacNaughton W.K. (2017). Signaling pathways induced by serine proteases to increase intestinal epithelial barrier function. PLoS ONE, 12(7), e0180259.
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8

Ussing Chamber Measurements of Bronchial Epithelia

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Snapwell supports carrying differentiated bronchial epithelia were mounted in Ussing-like vertical chambers (EM-CSYS-8, Physiologic Instruments). Both apical and basolateral chambers were filled with 5 mL of a Ringer bicarbonate solution containing (in mM): 126 NaCl, 0.38 KH2PO4, 2.13 K2HPO4, 1 CaCl2, 1 MgSO4, 24 NaHCO3, 10 glucose, and phenol red. Solution on both sides were bubbled with 5% CO2/95% air and kept at 37°C. The transepithelial voltage was clamped at 0 mV with an 8-channel voltage-clamp amplifier (VCC MC8, Physiologic Instruments) connected to apical and basolateral chambers via Ag/AgCl electrodes and agar bridges (1M KCl in 2% agar). The resulting short-circuited current from each channel was recorded with the Acquire & Analize 2.3 software (Physiologic Instruments).
Guidone D., Buccirossi M., Scudieri P., Genovese M., Sarnataro S., De Cegli R., Cresta F., Terlizzi V., Planelles G., Crambert G., Sermet I, & Galietta L.J. (2022). Airway surface hyperviscosity and defective mucociliary transport by IL-17/TNF-α are corrected by β-adrenergic stimulus. JCI Insight, 7(22), e164944.
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9

Epithelial Ion Transport Measurements

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Epithelia were assayed in Ussing chambers (Physiologic Instruments) with 3 M KCl agar bridges connected to amplifiers (VCC-MC8, Physiologic Instruments) recording open-circuit transepithelial voltage (Vt). A 5 μA bipolar current pulse was applied across the epithelium periodically. The current-induced change in Vt was used to calculate the transepithelial conductance (Gt). Data were acquired with Acquire & Analyze software (version 2.3.8, Physiologic Instruments). Dilution potentials were generated by perfusing dilutions of the dominant ionic species (e.g., NaCl) into the apical chamber. After the experiment, cells were lysed by distilled water and electrode drift was assessed in the original bilateral solution. Junction potentials induced in the voltage probes by ionic dilutions were then assessed without an epithelium and subtracted from obtained dilution potentials.
Thornell I.M., Rehman T., Pezzulo A.A, & Welsh M.J. (2020). Paracellular bicarbonate flux across human cystic fibrosis airway epithelia tempers changes in airway surface liquid pH. The Journal of Physiology, 598(19), 4307-4320.
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10

Ussing Chamber Assay for CFTR Modulation

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Differentiated hNECs were mounted in circulating Ussing chambers (VCC MC8; Physiologic Instruments, San Diego, CA). Short-circuit current (Isc, µA/cm2) was measured under asymmetric chloride (Cl−) Ringer's buffer (bicarbonate free) as previously described (47 (link)). After recording baseline current for 30 min, cells were sequentially treated with pharmacological compounds: 100 μM apical amiloride to inhibit epithelial sodium channels, 10 μM apical VX-770 (IVA) or 0.01% DMSO (vehicle) to potentiate CFTR-activated currents, 10 μM basal forskolin induce cAMP activation of CFTR, 30 μM apical CFTRInh−172 to inhibit CFTR-specific currents and 100 μM apical ATP to activate calcium-activated chloride currents. Data recordings were obtained using Acquire and Analysis 2.3 software (Physiologic Instruments, San Diego, CA). Isc in response to forskolin alone (no modulator treatment) was considered as baseline activity (ΔIsc−Fsk). Cumulative changes of Isc in response to forskolin and CFTR modulator were used as the measure of total CFTR-activated currents.
Allan K.M., Astore M.A., Fawcett L.K., Wong S.L., Chen P.C., Griffith R., Jaffe A., Kuyucak S, & Waters S.A. (2022). S945L-CFTR molecular dynamics, functional characterization and tezacaftor/ivacaftor efficacy in vivo and in vitro in matched pediatric patient-derived cell models. Frontiers in Pediatrics, 10, 1062766.
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