External Excitation of One-Dimensional Patterned Neuronal Cultures
Transcription
Take a coverslip containing a 1D patterned neuronal culture.
The neurons are loaded with a calcium indicator dye, which fluoresces upon calcium binding.
Transfer the coverslip to a fluorescence microscope imaging chamber containing an extracellular recording solution.
Assemble a holder with two parallel wire electrodes separated by a distance.
Invert the holder and position it above the neuronal culture with the electrodes immersed in the recording solution.
Connect the electrodes to an amplifier. Apply a non-DC pulsed bipolar square wave to generate a uniform uni-directional electric field across the neurons without causing injury.
The generated electric field excites the neurons, triggering an action potential.
The action potential travels along the axon, activating the voltage-gated calcium channels and causing calcium influx into the neuron terminal. Further, neurotransmitters are released into the synapse, facilitating synaptic transmission.
Following calcium ion influx, these ions bind to the intracellular dye and fluoresce, indicating neuronal activity in response to electrical stimulation.
