Initiating Rapid Neurite Extension and Formation Functional Neuronal Connections

Begin by assembling a microfluidic device containing microchannels within a dish coated with lysine.

Seed embryonic neurons with a growth medium into the microfluidic wells, allowing them to flow through the microchannels.

Incubate to facilitate neuronal attachment and growth, forming physically isolated neuronal populations.

Disassemble the setup and place the dish in a micromanipulator chamber with a saline flow to maintain cell viability.

Introduce lysine-coated beads near the neurons, enabling the neurites or cell projections to attach and form bead-neurite complexes.

Using a micromanipulator, apply suction to engage a bead-neurite complex from one channel.

Pull this complex to induce mechanical tension, leading to rapid neurite extension.

Guide this toward another bead-neuron complex and hold it with a second micromanipulator.

Keep both beads in place to facilitate the formation of a neuritic connection.

Release the suction and replace the saline with a culture medium.

Incubate to establish functional connections between neurons, promoting communication through neurotransmitters.