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Cytoview mea plate

Manufactured by Axion Biosystems
Sourced in United States

The CytoView MEA plates are a versatile tool for recording and analyzing the electrical activity of cells in vitro. These plates are designed to provide a high-quality interface for monitoring the electrophysiological responses of various cell types, including neurons, cardiomyocytes, and other excitable cells. The plates feature a multi-electrode array (MEA) that allows for the simultaneous recording of electrical signals from multiple points within the cell culture, enabling researchers to gain insights into the collective behavior and network dynamics of the cells under study.

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14 protocols using cytoview mea plate

1

Functional Analysis of Induced Sensory Neurons

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Accutase (StemCell Technology)-dissociated neurons were seeded at a density of 5 to 10 x 104 cells per well to fully cover all 16 embedded electrodes in a 24-well CytoView MEA plate (Axion Biosystems, Atlanta, GA) precoated with 0.1% polyethleneimine (PEI) and 300 μg/mL of Matrigel. iSNs were then cultured for at least 2 weeks to from the functional networks before analysis. Real-time well-wide neuronal spontaneous firings with a spike detection criterion of >6 STDs were recorded with MaestroEdge (Axion Biosystems) for 3 to 5 minutes at 37°C. Data were analyzed with AXIS Navigator and AXIS Neural Metric Tool. Only the cells in a well with at least 7 active electrodes were used for analysis, and the firing rate based on active electrodes was presented as the weighted mean firing rate. In the experiments with compound treatment, after the baseline firings were recorded, iSNs were then stimulated with 1 µM capsaicin (TRPV1 agonist), 100 µM AITC (TRPA1 agonist), or 100 µM menthol (TRPM8 agonist) (Sigma Aldrich, St. Louis, MO). The evoked neuronal activity was immediately recorded. Recovery for at least 24 hours is required between each treatment. In the study with Activin A stimulation, iSNs were incubated with Activin A (50ng/mL, R&D) for 30 minutes. All iSNs were allowed to recover for at least 24 hours before the next treatment.
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2

Cortical i3Neuron Electrophysiology on MEA

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Cortical i3Neurons were plated on a CytoView MEA plate (Axion Biosystems) at day 3 of differentiation. MEA recordings were performed on a Maestro Edge (Axion Biosystems) every 3-4 days during differentiation. Recordings on the MEA were performed prior to media changes. Neurons were removed from the incubator and were allowed to equilibrate in the recording chamber for 15 minutes prior to the start of recording. Neurons were recorded for 15 minutes, and the total number of spikes over the whole recording was reported.
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3

Multimodal Stimulation of Induced Neurons

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Accutase (StemCell Technology)-dissociated neurons were seeded at a density of 5–10 × 104 cells per well to fully cover all 16 embedded electrodes in a 24-well CytoView MEA plate (Axion Biosystems) precoated with 0.1% polyethleneimine (PEI) and 300 μg/ml of Matrigel. iSNs were then cultured for at least 2 weeks to from the functional networks before analysis. Real-time well-wide neuronal spontaneous firings with a spike detection criterion of >6 STDs were recorded with MaestroEdge (Axion Biosystems) for 3–5 minutes at 37 °C. Data were analyzed with AXIS Navigator and AXIS Neural Metric Tool. Only the cells in a well with at least 7 active electrodes were used for analysis, and firing rate based on active electrodes was presented as weighted mean firing rate. In the experiments with compound treatment, after the baseline firings were recorded, iSNs were then stimulated with 1 μM capsaicin (TRPV1 agonist), 100 μM AITC (TRPA1 agonist), or 100 μM menthol (TRPM8 agonist) (Sigma). The evoked neuronal activity was immediately recorded. Recovery for at least 24 hours is required between each treatment. In the study with Activin A stimulation, iSNs were incubated with Activin A (50ng/ml, R&D) for 30 mins. All iSNs were allowed to recover for at least 24 hours before the next treatment.
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4

Multielectrode Array Astrocyte-Neuron Co-culture

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Multielectrode array 24-well plates containing 16 electrodes per well were obtained from Axion Biosystems (CytoView MEA plate). The electrodes were arranged in a 4x4 grid 350 μm apart, with each electrode diameter being 50 μm. The electrode recording area was 1.1 mm x 1.1 mm. Plates were sterile upon arrival and prepared for cell adhesion by treating the electrode field with 0.1 mg/mL Poly-D-Lysine at 37° C for 4.5 hours. The wells were washed with water three times and dried in a biosafety cabinet overnight. 20 μg/mL laminin droplets were added to each field and incubated at 37° C for at least one hour. A master mix of neural progenitor cells and astrocytes was made (90,000 NPCs: 45,000 astrocytes per well) and plated in 12 μL drops on each electrode field. The cell culture droplets were incubated at 37° C for one hour, followed by adding 1 mL cortical media to fill the well. Water was added to the surrounding reservoirs to prevent media evaporation. Media changes occurred every 2-4 days by performing half media changes. Cells matured for about one month.
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5

Human iPSC-CM Irradiation Protocol

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Human iPSC-CMs (Cardiosight-S; NEXEL, Seoul, Korea) were used for MEA and real-time quantitative polymerase chain reaction (qPCR). The cells were prepared according to the manufacturer’s instruction. Briefly, 3 × 104 cells were seeded onto each well coated with fibronectin in the CytoView MEA plate (Axion BioSystems, Atlanta, GA, USA). After 1 h of incubation, Cardiosight-S maintenance medium was added, which was replaced every 2 days. Spontaneous beating was observed within 3 days of incubation at 37 °C and 5% CO2. Irradiation was performed 2 weeks after seeding.
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6

Multielectrode Array for Neuronal Activity

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A CytoView MEA plate (M384-tMEA-6B, Axion Biosystems) was used for all in vitro electrophysiology recordings. Each well of the MEA plate consists of 64 PEDOT electrodes (100 μm diameter, 300 μm inter-electrode spacing) arranged in an 8 × 8 grid at the center of each well. Daily recordings were performed with a Maestro Pro (Axion Biosystems) at the same time each day, from days in vitro (DIV) 25–49. The cultures were allowed to equilibrate for at least 15 min prior to each recording and incubated during the recording to ensure stable activity.
Activity was captured using AxIS 2.4 software (Axion Biosystems). Spike detection was performed on the raw data with the same software using the adaptive threshold method with a threshold of ±7 standard deviations from the median of the signal.
For the excitation-to-inhibition ratio (E/I) disruption assay, γ-aminobutyric acid (GABA; A5835, Sigma-Aldrich) was diluted in DPBS−/− and added immediately preceding recording on DIV 50.
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7

Analyzing Neuronal Activity in Mouse Cortical Neurons

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Mouse cortical neurons were plated in 48‐well CytoView MEA plates (Axion BioSystems) at a density of 160,000 cells per 10 μl containing 20 μg/ml laminin and transduced as above. Media was changed at DIV 12 and baseline spontaneous activity was recorded at DIV 14 for 15 min using the Maestro Pro MEA System (Axion BioSystems). Compounds or vehicle were added after baseline reading and recordings were repeated 48 h after compound addition. Extracellular voltage was recorded at each electrode with a sampling rate of 12.5 kHz. Spikes were identified from the raw signals using a detection threshold set independently for each electrode of ± 6x the standard deviation of the noise (AxIS Navigator software, Axion Biosystems). Electrodes with at least five spikes/min were considered active electrodes. The Neural Metric Tool (Axion BioSystems) was used for analysis of neuronal firing properties.
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8

Cardiomyocyte MEA Recording and CYP Treatment

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MEA recording in cardiomyocytes was performed as described previously (23 (link)). In brief, 2 × 104 cells were plated on CytoView MEA plates (Axion Biosystems, USA) pre-coated with 5% matrigel, followed by treatment with CYP at different concentrations (0 and 500 μmoL/L). The experimental data were acquired using a Maestro EDGE (Axion Biosystems, USA) according to the MEA operation manual.
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9

Neuronal Electrophysiology of Cultured Cells

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24 well Cytoview MEA plates (Axion Biosystems, Cat. No. M384-tMEA-24W) were coated with 0.1% PEI (MilliporeSigma, Cat. No. 03880) dissolved in 1X Borate Buffer (10mM Boric Acid, 2.5mM Sodium Tetraborate, 7.5mM NaCl, adjusted to pH 8.4) overnight at 37°C, washed with water, then dried at room temperature. Wells were then coated with BrainPhys media and 15 μg/mL Laminin. Cells were then seeded at a density of 200,000 cells per well on day 0 of differentiation. Measurements were taken using the Maestro Edge multiwell microelectrode array (MEA) and Impedance system (Axion Biosystems) at 3 × 5-minute intervals with the Maestro MEA platform software (Axion Biosystems) with the “Neural” default setting after 14 days in culture.
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10

Evaluating SARS-CoV-2 effects on hPSC-CMs

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CytoView MEA plates (Axion BioSystems) were coated with Matrigel and hPSC-CMs were plated as described previously (Bertero et al., 2019b (link)). SARS-CoV-2 effects on hPSC-CMs electrophysiology were recorded using the Maestro Pro system and analyzed with Cardiac Analysis Software v.3.1.8 (all from Axion BioSystems).
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