recording electrophysiological activity of the neuronal network in a neuron-astrocyte co-culture

Recording Electrophysiological Activity of the Neuronal Network in a Neuron-Astrocyte Co-culture

On the day of plating or before, start coating the 24-well MEA plates by first diluting PLO in water or PBS. Then, add 15 to 20 microliters of PLO to each well, forming a droplet on the center of the well, covering the electrode area, ensuring not to damage electrodes with the pipette tip.
Add water to the compartments&#39; surrounding wells, ensuring sufficient humidity, and incubate PLO at 37 degrees Celsius for 1 to 2 hours. Then, using a plastic micropipette tip, aspirate as much PLO as possible without touching the electrodes. Next, wash the well with 250 microliters of water, three times.
Using a pipette tip, remove as much water as possible and let the surface dry with the lid removed. Next, add 15 to 20 microliters of diluted laminin to cover each electrode array. After adding water to the humidity compartments and replacing the lid, incubate at 37 degrees Celsius for a minimum of two hours up to overnight.
On the day of plating, after rinsing the motor neurons and astrocyte cultures once with PBS, add 0.05% trypsin and incubate at 37 degrees Celsius for about 5 minutes to lift cells. Collect cells into a 15-milliliter conical tube, containing trypsin inhibitor, and wash plates with medium or base to ensure that all the cells are collected.
Pellet down the cells by centrifugation, and then using a 1,000-microliter pipette, resuspend motor neurons and astrocytes with a co-culture medium containing 20 micromolar of ROCK inhibitor to generate 1 milliliter of a single-cell suspension. Using a hemocytometer, count the motor neurons and astrocytes, and while counting, cap the cell suspensions and place them in a styrofoam rack at 4 degrees Celsius.
Centrifuge 1 milliliter of both cell suspensions at 300 times g for 5 minutes. Calculate the desired volume, and re-suspend the pellets. Combine the required volume of neuron and astrocyte suspensions in equal ratios, and mix gently by pipetting twice to combine thoroughly.
Then, remove laminin from each well of the plate and transfer 10 microliters of the final combined cell suspension to each well, forming a small droplet covering the electrode array. Incubate the plates with an undisturbed cell droplet at 37 degrees Celsius for 20 to 30 minutes to form initial attachments on the plates.
Then, pipette 250 microliters of warm co-culture media containing ROCK inhibitor down on the wall of each well, and pipette the same volume on the opposite wall of each well, and incubate the plate at 37 degrees Celsius for 24 to 36 hours. The next day, examine the plates for debris or dead cells, and if required, exchange the medium with a fresh co-culture medium containing ROCK inhibitor.
Otherwise, perform half-medium exchanges twice a week using a co-culture medium without a ROCK inhibitor, starting on co-culture day 2. To perform recording, start as soon as possible after co-culture day 1 by setting the temperature to 37 degrees Celsius and carbon dioxide at 5%. Then, transfer plates to the recording machine, and equilibrate for at least five minutes before recording. Record baseline activity every other day or weekly over 1 to 15 minutes, depending on the experimental design.

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1. Co-culturing MN and spinal cord astrocytes in multi-electrode array plates
<ol>
	<li>Differentiate and plate the motor neurons and astrocytes when they are ready to be used on the same day and aged to day in vitro (DIV) 60 for the motor neurons and DIV 90 for the astrocytes.</li>
	<li>Coat the MEA plates on the day before or on the day of plating as follows if working with 24-well plastic plates (Table of Materials).</li>
	<li>Dilute polyornithine (PLO) in water or phosphate-buffered saline (PBS) to 100 &#181;g/mL.
	<ol>
		<li>Add 15-20 &#181;L to each well (dependent on pipette comfort level), forming a droplet on the center of the well covering the area of the electrodes and surrounding area but not the entirety of the well.</li>
		<li>Take care not to damage electrodes with the pipette tip. Be consistent with volume from well to well to ensure coverage of the same surface area in each well.</li>
		<li>Incubate PLO at 37 &#176;C for a minimum of 1 h (preferably 2 h). 
		NOTE: The small volumes will dry up if there is not sufficient humidity in the plates. Add water to the compartments surrounding wells to ensure sufficient humidity throughout the course of coating and recordings.</li>
	</ol>
	</li>
	<li>Aspirate as much PLO as possible using a plastic micropipette tip. Take care not to touch the electrodes. Wash with 250 &#181;L of water three times. If using a vacuum aspirator for washes, do not allow the tip near the electrode array. After the third wash, remove as much water as possible, using a pipette tip as necessary. Let the surface dry under the cell culture hood with the lid removed.</li>
	<li>Once plate surfaces are dry, add laminin diluted to 10 &#181;g/mL in PBS. Use 15-20 &#181;L to cover each electrode array. Add water to the humidity compartments, replace the lid, and return the plate to 37 &#176;C incubation for a minimum of 2 h up to overnight.</li>
	<li>On the day of plating, rinse the motor neuron (MN) and astrocyte cultures once with PBS and add trypsin 0.05% at 37 &#176;C to lift cells (5 min). Collect into a 15 mL conical tube containing trypsin inhibitor and wash plates with medium or base to ensure that all the cells are collected. Centrifuge at 300 x g for 5 min and resuspend with a 1000 &#181;L pipette to generate 1 mL of a single cell suspension. When re-suspending MN and astrocytes, switch to the co-culture medium (Table 1), with the addition of 20 &#181;M Rho-associated coiled-coil forming protein serine/threonine kinase inhibitor (ROCK-I).</li>
	<li>Count the MN and astrocytes in parallel using a hemocytometer. While counting and making calculations, cap the cell suspensions and place them in a Styrofoam rack at 4 &#176;C.</li>
	<li>Calculate the volume required to resuspend the cultures to a concentration of 5 x 104 cells per 5 &#181;L for motor neurons and 2.5 x 104 cells per 5 &#181;L for astrocytes. Centrifuge the 1 mL cell suspensions at 300 x g for 5 min and resuspend in the calculated volume.</li>
	<li>Calculate the number of desired wells to be seeded and multiply it by 5 &#181;L of each cell suspension. Combine the required volume of neuron and astrocyte suspensions at a 1:1 ratio and mix by pipetting until thoroughly combined (usually twice) but avoid being too aggressive.</li>
	<li>Remove the laminin from each well of the MEA plate using a pipette tip. Transfer 10 &#181;L of the final combined cell suspension to each well, forming a small droplet covering the electrode array to provide a cell density of 5 x 104 MN and 2.5 x 104 astrocytes per well. 
	NOTE: High cell density in the combined suspension requires frequent resuspension in between wells during the seeding step to ensure accurate and consistent cell counts.</li>
	<li>Return the plates to the incubator for 20-30 min. Take care not to disturb the cell droplet and allow the cells to form initial attachments on the plates.</li>
	<li>After 20-30 min, add warm co-culture media + ROCK-I to each well by pipetting 250 &#181;L down on the wall of each well, followed by an additional 250 &#181;L down the wall of the same well on the opposite side. Return the plate to the incubator.</li>
	<li>Examine plates the day after seeding (Co-culture Day 1). If there is significant cell debris or dead cells, exchange the medium with a fresh co-culture medium containing ROCK-I. Otherwise, change the medium on the second day after seeding (Co-Culture Day 2) to the co-culture medium without ROCK-I. Perform half-medium exchanges (aspirate 50% and add 60% of the final volume to account for evaporation) twice a week.</li>
	<li>If using MEA systems with single well 30 mm glass plates (Table of Materials), the plate preparation may take 2 or more days. Follow the steps below to perform this.</li>
	<li>Lift the cells and cell debris from the previous MEA cultures with 0.05% Trypsin for 5-15 min.</li>
	<li>Aspirate trypsin and rinse three times with water.</li>
	<li>Sterilize by adding 70% ethanol and incubate in the hood for 10 min. Aspirate the ethanol, and then rinse with water three times.</li>
	<li>Apply 1% anionic detergent with protease enzyme in water. Cover the plates with a lid and wrap with thermoplastic film and aluminum foil, and then leave them on a rocker at room temperature overnight.</li>
	<li>Aspirate the detergent, and then rinse three times with water.</li>
	<li>If planning to store the plates, add a sufficient volume of water. Cover the plates with a lid and wrap them with thermoplastic film and aluminum foil and leave them in the refrigerator until next use.</li>
	<li>If planning to coat, let the surface dry under the cell culture hood.</li>
	<li>If extra sterilization is needed (e.g., previous infection or non-recent use), at this point only, expose MEA plates to ultraviolet (UV) light under the hood for 30-60 min. 
	NOTE: Avoiding frequent UV light will prevent damage to the electrodes. The use of UV light when plates have serum on them will result in nonfunctional electrodes, which is why it should only be used on cleaned plates.</li>
	<li>When the plates are completely dry, proceed with plasma cleaning for 1 min to charge the glass surface of MEA plates and enhance coating effectiveness. Plasma-clean at least once every 4-5 cycles of MEA plate cleaning-plating cycles.
	<ol>
		<li>As an alternative, when plasma cleaning is not feasible or not warranted, add a sufficient volume of fetal bovine serum (FBS)-containing medium to cover the MEA plates&#39; surface and return them to the incubator overnight.</li>
	</ol>
	</li>
	<li>Use PLO (polyornithine)/Laminin coating. Add PLO solution (100 &#181;g in PBS) immediately after plasma cleaning or aspirating and rinsing out the FBS-containing medium.</li>
	<li>Plate the cells as above at the following densities: 1 x 105 human pluripotent stem cell-derived astrocytes (hiPSC-A) / plate and 5 x 105 human pluripotent stem cell-derived motor neurons (hiPSC-MN)/plate.</li>
</ol>
2. Multi-electrode array recording
<ol>
	<li>Extract the raw voltage data at a 12.5 kHz sampling frequency, using a Butterworth filter with a 200 Hz high pass and 3 kHz low pass filter. Set spike recognition as instantaneous time points of voltages &#8805;6 standard deviations from baseline. Identify bursts as an activity with &#62;5 spikes in 100 ms. Define network activity when over 35% of total active electrodes fire within 100 ms with a minimum of 50 spikes per network burst.</li>
	<li>Start recording as soon as possible after Co-Culture (CC) Day 1. 
	NOTE: Electrical activity will be rarely noted before CC Day 3.</li>
	<li>Plate parallel cultures at similar densities and replicates in the appropriate culture plates or coverslips for biochemical assays, immunocytochemistry (ICC), or quantitative PCR (qPCR).</li>
	<li>Perform recordings with the temperature set to 37 &#176;C and CO2 at 5%. Transfer plates to the machine and allow them to equilibrate for at least 5 min prior to recording.</li>
	<li>Record baseline activity either every other day or weekly, over 1-15 min depending on the experimental design. Do not record for at least 1 h after a medium exchange, and ideally wait for 24 h after every media exchange.</li>
	<li>To prevent contamination, change the medium (i.e., half medium exchange as in step 1.14) after any recording in which the lid of the sterile plate has to be opened outside of the sterile hood (i.e., the application of drug compounds). Perform complete medium exchanges and washes to wash out drugs if plates are going to continue to be used beyond drug treatment.</li>
	<li>For analysis purposes, use at least three technical and biological replicates for each condition. Express electrophysiological data as means of n &#8805; 3 replicates.</li>
</ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>10 cm sterile culture plates</td>
			<td>Falcon</td>
			<td>353003</td>
			<td></td>
		</tr>
		<tr>
			<td>25 cm2 sterile culture flasks</td>
			<td>Falcon</td>
			<td>353136</td>
			<td></td>
		</tr>
		<tr>
			<td>500 mL 0.2 &#181;m CA Filter System</td>
			<td>Corning</td>
			<td>430769</td>
			<td></td>
		</tr>
		<tr>
			<td>5 mL pipette</td>
			<td>Falcon</td>
			<td>357543</td>
			<td></td>
		</tr>
		<tr>
			<td>6 well sterile culture plates</td>
			<td>Falcon</td>
			<td>3046</td>
			<td></td>
		</tr>
		<tr>
			<td>Axion CytoView MEA 24 plates (M384-tMEA-24W)</td>
			<td>Axion</td>
			<td>OPT-24</td>
			<td></td>
		</tr>
		<tr>
			<td>Axion Edge MEA platform</td>
			<td>Axion</td>
			<td>Maestro Edge</td>
			<td></td>
		</tr>
		<tr>
			<td>Basement Membrane matrix - Matrigel</td>
			<td>Corning</td>
			<td>354277</td>
			<td>Details in the protocol</td>
		</tr>
		<tr>
			<td>Benchtop microscope (sterile under cell culture hood)</td>
			<td>Zeiss</td>
			<td>415510-1100-000</td>
			<td>Primo Vert</td>
		</tr>
		<tr>
			<td>Ciliary neurotrophic factor (CNTF)</td>
			<td>Peprotech</td>
			<td>450-13</td>
			<td>Dissolve 100 &#181;g in 1mL of sterile PBS, and then add 9 mL of sterile 0.1% BSA-PBS to 10 &#181;g/mL. Aliquot and freeze at -80 &#186;C. Dilute at 1: 1000 for use. (working concentration 10 ng/mL).</td>
		</tr>
		<tr>
			<td>CO2 tanks and regulator for Axion Edge</td>
			<td>AirGas/Harris</td>
			<td>9296NC</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM/F12</td>
			<td>Thermofisher</td>
			<td>113300</td>
			<td>Working concentration 1x</td>
		</tr>
		<tr>
			<td>Essential 8 Medium + Essential 8 Supplement</td>
			<td>Thermofisher</td>
			<td>A1517001</td>
			<td>Combine 10 mL of Essential 8 (10x)&#160; supplement with 500 mL of Essential 8 Growth Medium (1x)</td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum (FBS)</td>
			<td>Thermofisher</td>
			<td>16140071</td>
			<td>Working concentration 1x</td>
		</tr>
		<tr>
			<td>Hemocytometer</td>
			<td>Election Microscopy Sciences</td>
			<td>63510-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Humidity controlled Cell culture incubator</td>
			<td>ThermoFisher</td>
			<td>370</td>
			<td>set to 37 &#186;C, 5 % CO2</td>
		</tr>
		<tr>
			<td>Laminin</td>
			<td>Thermofisher</td>
			<td>23017-015</td>
			<td>Stock solution 1 mg/mL, working concentration 10 &#181;g/mL (coating), 1 &#181;g/mL (cell media)</td>
		</tr>
		<tr>
			<td>L-Glutamine</td>
			<td>Thermofisher</td>
			<td>25030</td>
			<td>Dissolve 2 mg of LDN into 500 &#181;L of Chloroform to get 10 mM stock. Aliquot this and freeze at -80 &#186;C. For using, dilute the stock 1 to 10 into DMSO [to 1 mM] first, then dilute 1:5000 of 1 mM into the desired media to get 0.2 &#181;M working solution</td>
		</tr>
		<tr>
			<td>MEA glass plates</td>
			<td>MultiChannel Systems</td>
			<td>60MEA200/30iR-Ti-gr</td>
			<td>Working concentration 100x</td>
		</tr>
		<tr>
			<td>Multichannel Pipet P200</td>
			<td>Gilson</td>
			<td>PJ22224</td>
			<td></td>
		</tr>
		<tr>
			<td>Neurobasal</td>
			<td>Thermofisher</td>
			<td>21103049</td>
			<td></td>
		</tr>
		<tr>
			<td>Non-Essential Amino Acids (NEAA)</td>
			<td>Thermofisher</td>
			<td>11140050</td>
			<td>Working concentration 1x</td>
		</tr>
		<tr>
			<td>Pencillin/Streptomycin</td>
			<td>Thermofisher</td>
			<td>15140122</td>
			<td>Working concentration 100x</td>
		</tr>
		<tr>
			<td>Polyornithine (PLO)</td>
			<td>Sigma-Aldrich</td>
			<td>P3655</td>
			<td>Working concentration 100x</td>
		</tr>
		<tr>
			<td>Potassium chloride (KCl)</td>
			<td>NA</td>
			<td>NA</td>
			<td>Dissolve 100 mg in 1 mL of ddW to get 100 mg/mL stock solution. Aliquot the stock in 100 &#181;L tubes. (working concentration 100 &#181;g/mL)</td>
		</tr>
		<tr>
			<td>Purmorphamine (PMN)</td>
			<td>Millipore-Sigma</td>
			<td>540223</td>
			<td>Working concentration100 mM</td>
		</tr>
		<tr>
			<td>ROCK-Inhibitor</td>
			<td>Peprotech</td>
			<td>1293823</td>
			<td>Centrifuge briefly before reconstitution. Dissolve 100 &#181;g in 1 mL of PBS to 100 &#181;g/mL, and then add 9 mL of sterile 0.1% BSA-PBS to 10 &#181;g/mL. Aliquot and freeze at -80 &#186;C. Dilute at 1: 1000 for use. (working concentration 10 ng/mL).</td>
		</tr>
		<tr>
			<td>Sterile cell culture hoods</td>
			<td>Baker Company</td>
			<td>SG-600</td>
			<td>Dissolve 5 mg in 1480 &#181;L of dH2O to get 10 mM stock, aliquot and freeze at -80 &#186;C. Dilute at 1: 500 for use. (working concentration 20 &#181;M).</td>
		</tr>
		<tr>
			<td>Supplement B - B27 Supplement</td>
			<td>Thermofisher</td>
			<td>21985023</td>
			<td></td>
		</tr>
		<tr>
			<td>Table top cell culture centrifuge</td>
			<td>ThermoFisher</td>
			<td>75004261</td>
			<td>Working concentration 50x</td>
		</tr>
		<tr>
			<td>Thermoplastic film - Parafilm</td>
			<td>PARAFILM</td>
			<td>P7793</td>
			<td>Sorvall Legend X1R</td>
		</tr>
		<tr>
			<td>Trypsin-EDTA (0.05%)</td>
			<td>Thermofisher</td>
			<td>2530054</td>
			<td>Working concentration 1x</td>
		</tr>
		<tr>
			<td>Waterbath</td>
			<td>ThermoFisher</td>
			<td>2332</td>
			<td>Isotemp</td>
		</tr>
	</tbody>
</table>

Source: Taga, A., et al. <a href="https://app.jove.com/v/62726">Establishment of an Electrophysiological Platform for Modeling ALS with Regionally-Specific Human Pluripotent Stem Cell-Derived Astrocytes and Neurons.</a> J. Vis. Exp. (2021).
This video demonstrates establishing a neuron-astrocyte co-culture for recording neuronal network activity. Upon establishing the co-culture of motor neurons and astrocytes on a multi-electrode array (MEA) plate, the synchronous rhythmic electrical activity of the neuronal network was recorded extracellularly using the MEA.

1. Co-culturing MN and spinal cord astrocytes in multi-electrode array plates

	Differentiate and plate the motor neurons and astrocytes when they are ...

Begin with a multi-electrode array, or MEA, plate. Each well contains a centrally positioned array of electrodes.
The array is coated with a culture substrate to facilitate cell adhesion.
Add a suspension of motor neurons and astrocytes as a droplet over the array.
Incubate to allow cell attachment, then add a warm co-culture medium supplemented with a small molecule that maintains cell viability.
During incubation, astrocytes secrete factors that promote neuronal maturation and synapse formation, resulting in a neuronal network.
Place the MEA in a recording device under physiological conditions to monitor neuronal activity.
Neurons transmit electrical signals, called action potentials, that travel along their axons.
Upon reaching the synapse, the signal triggers neurotransmitter release, propagating the signal to the next neuron.
Electrodes within each well extracellularly record the signals generated by synaptically connected neurons.
The detection of synchronized rhythmic electrical signals confirms the establishment of a neuronal network.

22 次观看. 记录神经元-星形胶质细胞共培养物中神经元网络的电生理活性

视频: 记录神经元-星形胶质细胞共培养物中神经元网络的电生理活性 - 实验

Electrophysiological Investigations of Retinogeniculate and Corticogeniculate Synapse Function

The lateral geniculate nucleus is the first relay station for the visual information. Relay neurons of this thalamic nucleus integrate input from retinal ganglion cells and project it to the visual cortex. In addition, relay neurons receive top-down excitation from the cortex. The two main excitatory inputs to the relay neurons differ in several aspects. Each relay neuron receives input from only a few retinogeniculate synapses, which are large terminals with many release sites. This is reflected by the comparably strong excitation, the relay neurons receive, from retinal ganglion cells. Corticogeniculate synapses, in contrast, are simpler with few release sites and weaker synaptic strength. The two synapses also differ in their synaptic short-term plasticity. Retinogeniculate synapses have a high release probability and consequently display a short-term depression. In contrast, corticogeniculate synapses have a low release probability. Corticogeniculate fibers traverse the reticular thalamic nuclei before entering the lateral geniculate nucleus. The different locations of the reticular thalamic nucleus (rostrally from the lateral geniculate nucleus) and optic tract (ventro-laterally from the lateral geniculate nucleus) allow stimulating corticogeniculate or retinogeniculate synapses separately with extracellular stimulation electrodes. This makes the lateral geniculate nucleus an ideal brain area where two excitatory synapses with very different properties impinging onto the same cell type, can be studied simultaneously. Here, we describe a method to investigate the recording from relay neurons and to perform detailed analysis of the retinogeniculate and corticogeniculate synapse function in acute brain slices. The article contains a step-by-step protocol for the generation of acute brain slices of the lateral geniculate nucleus and steps for recording activity from relay neurons by stimulating the optic tract and corticogeniculate fibers separately.

Here, we present protocols for the preparation of acute brain slices containing the lateral geniculate nucleus and the electrophysiological investigation of retinogeniculate and corticogeniculate synapses function. This protocol provides an efficient way to study synapses with the high-release and low-release probability in the same acute brain slices.

视网膜原酸和皮质化突触功能的电生理学研究

The lateral geniculate nucleus is the first relay station for the visual information. Relay neurons of this thalamic nucleus integrate input from retinal ...

科研

JoVE Journal

神经科学

Preparation of Acute Human Hippocampal Slices for Electrophysiological Recordings

Epilepsy affects about 1% of the world population and leads to a severe decrease in quality of life due to ongoing seizures as well as high risk for sudden death. Despite an abundance of available treatment options, about 30% of patients are drug-resistant. Several novel therapeutics have been developed using animal models, though the rate of drug-resistant patients remains unaltered. One of probable reasons is the lack of translation between rodent models and humans, such as a weak representation of human pharmacoresistance in animal models. Resected human brain tissue as a preclinical evaluation tool has the advantage to bridge this translational gap. Described here is a method for high quality preparation of human hippocampal brain slices and subsequent stable induction of epileptiform activity. The protocol describes the induction of burst activity during application of 8 mM KCl and 4-aminopyridin. This activity is sensitive to established AED lacosamide or novel antiepileptic candidates, such as dimethylethanolamine (DMEA). In addition, the method describes induction of seizure-like events in CA1 of human hippocampal brain slices by reduction of extracellular Mg2+ and application of bicuculline, a GABAA receptor blocker. The experimental set-up can be used to screen potential antiepileptic substances for their effects on epileptiform activity. Furthermore, mechanisms of action postulated for specific compounds can be validated using this approach in human tissue (e.g., using patch-clamp recordings). To conclude, investigation of vital human brain tissue ex vivo (here, resected hippocampus from patients suffering from temporal lobe epilepsy) will improve the current knowledge of physiological and pathological mechanisms in the human brain.

The presented protocol describes the transport and preparation of resected human hippocampal tissue with the ultimate goal to use vital brain slices as a preclinical evaluation tool for potential antiepileptic substances.

为电子生理记录准备急性人类河马切片

Epilepsy affects about 1% of the world population and leads to a severe decrease in quality of life due to ongoing seizures as well as high risk for ...

Focused Ultrasound Neuromodulation of Human In Vitro Neural Cultures in Multi-Well Microelectrode Arrays

The neuromodulatory effects of focused ultrasound (FUS) have been demonstrated in animal models, and FUS has been used successfully to treat movement and psychiatric disorders in humans. However, despite the success of FUS, the mechanism underlying its effects&#160;on neurons remains poorly understood, making treatment optimization by tuning FUS parameters difficult. To address this gap in knowledge, we studied human neurons in vitro using neurons cultured from human-induced pluripotent stem cells (HiPSCs). Using HiPSCs allows for the study of human-specific neuronal behaviors in both physiologic and pathologic states. This report presents a protocol for using a high-throughput system that enables the monitoring and quantification of the neuromodulatory effects of FUS on HiPSC neurons. By varying the FUS parameters and manipulating the HiPSC neurons through pharmaceutical and genetic modifications, researchers can evaluate the neural responses and elucidate the neuro-modulatory effects of FUS on HiPSC neurons. This research could have significant implications for the development of safe and effective FUS-based therapies for a range of neurological and psychiatric disorders.

Focused Ultrasound Neuromodulation of Human In Vitro Neural Cultures in Multi-Well Microelectrode Arrays

Here, we present a protocol for using a high-throughput system that enables the monitoring and quantification of the neuromodulatory effects of focused ultrasound on human-induced pluripotent stem cell (HiPSC) neurons.

多孔微电极阵列中人 体外 神经培养物的聚焦超声神经调控

The neuromodulatory effects of focused ultrasound (FUS) have been demonstrated in animal models, and FUS has been used successfully to treat movement and ...

Recording Electrophysiological Activity of the Neuronal Network in a Neuron-Astrocyte Co-culture

成績單

Recording Electrophysiological Activity of the Neuronal Network in a Neuron-Astrocyte Co-culture