In this video we demonstrate the preparation of E18 Cortical Rat Neurons.
Understand the rationale for E18 cortical neuron isolation and microfluidic compartmentalization for axonal regeneration studies.
Sterilize dissection instruments, prepare enzymatic digestion solutions, and set up the microfluidic device and culture plates.
Incubate isolated cortical tissue in papain solution to dissociate cells from the tissue matrix.
Pipette the digested tissue through progressively smaller-bore pipette tips to separate individual neurons.
Plate dissociated neurons into the microfluidic device at appropriate density to establish axonal growth into separate channels.
Visualize axon and soma compartments in the microfluidic device using phase contrast or fluorescent markers.
Mechanically sever axons within the microfluidic device using a focused laser or micropipette to initiate regeneration studies.
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