Electrical activity of the human cardiomyocytes plated on patterned MEAs was recorded following 4–10 days in vitro (DIV) using a Multichannel Systems 60 channel amplifier (MEA 1040, Multichanel Systems). Prior to recording, the cells were allowed to equilibrate for 15 min in the lab atmosphere at 37°C. Temperature was maintained with a TC02 temperature controller (Multichannel Systems). The cells were stimulated using a STG 1002 stimulator (Multichannel Systems) by applying 1 ms wide bipolar square pulses of 500–700 mV amplitude at increasing frequencies ranging from 0.5 Hz to 2.5 Hz in 0.5 Hz increments. The recording medium was the same as the plating medium. For drug experiments, sotalol, (Sigma, cat#S0278), norepinephrine (Sigma, cat#A7257) or verapamil (Sigma, cat#381195) were added to the bathing medium in increasing concentrations of 10, 30, 100 and 300 μM – sotalol, 0.1, 0.3, 1 and 3 μM – norepinephrine, and 0.3, 1, 3 and 10 μM – verapamil. In all drug experiments, for control and each new drug concentration a total of 5 minutes of recordings were performed, the first 150 seconds with no stimulation, followed by 30 seconds of stimulation, and ended with no stimulation. The data was converted to pClamp (Axon Instruments) format using MC-Data Tool (Multichannel Systems) and analyzed with Clampfit (Axon Instruments) software and software written in Matlab.
Tc02 temperature controller
Manufactured by MultiSciences Biotech
The TC02 temperature controller is a laboratory instrument designed to precisely control and monitor the temperature of various experimental or process environments. It features a digital display, adjustable temperature setpoints, and integrated control algorithms to maintain the desired temperature within the specified range.
Automatically generated - may contain errors
Lab products found in correlation
Verapamil, by Merck Group (1 mentions)
Norepinephrine, by Merck Group (1 mentions)
Stg 1002 stimulator, by MultiSciences Biotech (1 mentions)
Mc datatool, by MultiSciences Biotech (1 mentions)
Pclamp, by Molecular Devices (1 mentions)
Clampfit, by Molecular Devices (1 mentions)
Sotalol, by Merck Group (1 mentions)
Mea2100 lite system, by MultiSciences Biotech (1 mentions)
Multi channel experimenter software, by MultiSciences Biotech (1 mentions)
Poly d lysine, by Merck Group (1 mentions)
Laminin, by Merck Group (1 mentions)
Labchart 8, by ADInstruments (1 mentions)
3 protocols using tc02 temperature controller
1
Electrophysiological Assessment of Cardiomyocytes on MEAs
2
Extracellular Recording of Primary Neurons on MEAs
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MEAs (Multi Channel Systems, 60MEA200/10iR-Ti) consist of 59 titanium nitride (TiN) working electrodes with a diameter of 10 μm with 200 μm spacing between electrodes and one internal reference electrode. Prior to recording, MEAs were treated with oxygen plasma for 30 seconds using a PX-500 Plasma System (March Instruments), 100 μg/mL of poly-D-lysine overnight (Sigma, P0899), and 10 μg/mL Laminin (Sigma, L2020) for 2h prior to culture of primary neurons. For recording, culture medium was removed and 1 mL of warmed MEA recording solution85 (link),86 (link) was added to each MEA. Each culture was allowed to equilibrate with the MEA solution for 5 minutes in the 37°C incubator before recording. Recordings were conducted with a MEA2100-Lite-System (Multi Channel Systems) that was maintained at 37°C using the TC02 temperature controller (Multi Channel Systems). The Multi Channel Experimenter software (Multi Channel Systems) was used to record the extracellular potential at each electrode for 5 minutes using a sampling rate of 20 kHz. After recording, MEA recording solution was removed and the conditioned cell culture medium was returned to the MEA.
3
Thermal Responsiveness of Sensory Neurons
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Utilizing TC02 temperature controller (Multi Channel Systems), thermal stimulation was conducted in MEAs plated with sensory neurons once every week starting from the third week of differentiation. Temperature was controlled via the software “TCX-Control Program”. Thermal assays include two protocols. The first protocol included two consecutive baseline recording blocks at 37 °C each for 3 minutes followed by a third block where incremental temperature increases were applied to the MEA (1 °C/30 seconds) from 37 °C to 45 °C and cooled back to 37 °C. The second protocol is similar to protocol 1 except in the third block of recording the MEA was maintained at 37 °C for one minute then suddenly heated to 45 °C for another minute and suddenly cooled back to 37 °C and recorded further for one minute. On some occasions, single units were discriminated in the recordings from each channel according to their relative amplitude and duration using Spike Histogram software (LabChart 8, ADInstruments). Graphs were generated using GraphPad Prism where data presented as scatter plot (mean) with error bars representing SEM.
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