Simultaneous Optical and Electrophysiological Monitoring of Neuronal Cells in Brain Slices
Transcrição
Start by perfusing an aerated artificial cerebrospinal fluid or aCSF over a mesh grid in a perfusion chamber.
Transfer a brain slice containing fluorescent protein-expressing neurons and anchor it with a platinum-wired device.
Rotate the mesh grid for alignment.
Lower the multichannel probe toward the slice.
Adjust the filter cube for fluorescent protein visualization.
Rotate the slice to align the probe.
Adjust the probe height below the tissue surface.
Now, perfuse the aCSF and focus on the fluorescently labeled neurons.
Using a high-power objective, focus on the tissue and adjust the excitation light.
Use an appropriate light filter to observe fluorescence in neurons.
Adjust the objective lens to create space for the patch pipette and apply positive pressure.
Next, position the pipette near the neuron to form a membrane dimple.
Finally, apply weak suction to create a membrane seal, followed by strong suction to break the membrane and obtain electrical recordings from the targeted neuron.
