Recording Neural Activity of Unfolded Hippocampal Tissue Using Penetrating Microelectrode Array
筆記録
In this procedure, fill one bottle with normal aCSF and another bottle with 4-AP aCSF. Bubble the solutions with 95% oxygen, 5% carbon dioxide from the beginning of the experiments. Use a trivalve connector to control which solution will be selected during an experiment.
Next, connect a vacuum tube to the outlet of the chamber to remove the solution into a dust container. Heat the pipeline before delivering the solution into the recording chamber and keep it at 35 degrees Celsius controlled temperature. After closing the inlet and outlet of the recording chamber, use a custom-made glass pipette dropper to transfer the unfolded hippocampus to the recording chamber.
Under the microscope, position the unfolded hippocampus with its obvious side facing down, CA3 area pointing away, and CA1 field pointing towards the researcher. Carefully aspirate away the solution in the chamber using a vacuum pipette from the edge of the recording chamber until the chamber is dried and the tissue lies on the array.
Then, carefully place a custom-made tissue anchor on top of the tissue to hold the unfolded hippocampus onto the array. Refill the recording chamber with a few drops of solution. Gradually open the inlet and outlet to adjust the flow rate to about two drops per second in the IV drip chamber.
Incubate the tissue in the recording chamber with normal aCSF for about one minute. Then, switch the solution supply to 4-AP dissolved aCSF, and adjust the flow rate properly. Incubate the tissue in 4-AP dissolved aCSF for about 5 to 10 minutes before recording.
