Isolating and Culturing Chick Embryonic Neurons on a Multi-Electrode Array

Take a fertilized chicken egg containing an early-stage embryo.

Sterilize the shell and carefully open it.

Remove the embryo. Dissect and transfer the head to a culture plate containing a buffer. Remove the eyeball.

Make an incision and remove the skin layers, exposing the forebrain and midbrain.

Remove the inner membranous layer and blood vessels. Dissect and transfer the forebrain to a culture plate with a buffer.

Cut the tissue into smaller fragments and transfer them to a conical tube. Once the fragments settle, remove the buffer.

Add a proteolytic enzyme solution. Incubate to digest the tissue's extracellular matrix and loosen neurons.

Remove enzyme-rich media, wash with neurobasal media, removing any remaining enzymes.

Add neurobasal media. Mechanically dissociate the tissue to obtain a neuronal suspension.

Seed these neurons on a multi-electrode array system coated with an extracellular matrix. Add neurobasal media and incubate.

Neurons attach and extend projections, forming a neuronal network.

On the day of neuron plating, remove a vial of extracellular matrix from the -20 degrees Celsius freezer. Spray with 70% ethanol and place on ice. Be sure to thaw out the extracellula