Inducing Axonal Varicosities with a Micromechanical Stimulus on Neurons
Transkript
Begin by securing a micropipette with attached tubing to a micromanipulator.
Attach the other end of the tubing to a syringe filled with a salt buffer via a connector with a turnable valve.
Set the syringe at a height that provides sufficient pressure to induce varicosities in axons.
Axonal varicosities are bead-like swellings that occur along the length of an axon.
Position the micropipette at the desired angle relative to the culture dish.
Use fluorescent light from the microscope to align the micropipette tip and lower it near the surface of the cell culture dish containing salt buffer.
Now, turn on the transmitted light and turn off the fluorescent filter.
Place a coverslip with neurons onto the dish.
First, focus on the micropipette and adjust its position.
Then, focus on the region containing the target neurons.
Open the valve to apply fluid pressure. This micromechanical stimulus induces varicosities in these neurons.
