Utilizing Multielectrode Arrays to Measure Synaptic Physiology in 3D Coculture Spheres

Take a sterile multielectrode array or MEA, a device that records the electrical activity of target cells.

Add a polymer solution to the MEA and incubate.

The polymer coats the surface of the MEA, thereby increasing the water-attracting property of the surface.

Remove the excess polymer and wash with deionized water.

Add extracellular matrix, or ECM, and incubate to allow it to coat the MEA. Remove the excess ECM.

Next, add the 3D coculture spheres comprising neurons and astrocytes in a suitable media.

Incubate to allow the spheres to adhere to the surface of the MEA.

Place the MEA on a temperature-controlled head stage of an MEA system and record the spontaneous electrical activity across the electrodes.

The measured electrical activity corresponds to the communication at the neural junctions within the sphere.

Begin by coating the surface of each multielectrode array or MEA with 1 millimeter of poly-ornithine to render the surface hydrophilic and incubate at 37 degrees Celsius for a minimum of four hours. Following the incubation, remove the poly-ornithine and wash the surface of each MEA with deionized water. Then, add 1 milliliter of extracellular matrix and return to the incubator for a minimum of four hours.

When ready, remove the extracellular matrix, then apply the spheres in 1.5 milliliters of medium onto the MEA surface, ensuring that the spheres are positioned on top of the electrode array. Allow to adhere for two days. To measure the electrical activity of neural spheres, place the MEA on a temperature-controlled head stage.