Week 6-7: Patch-Clamp Process
Welcome back! This post is a joint post for last week and this week.
At the University of Chicago, I conducted an experiment utilizing the patch-clamp technique to record the action potentials of a living neuron injected with 4-Aminopyridine. Prior to conducting the experiment, the Zebra Finch was paralyzed with isofluorane, and then the brain was carefully immersed in cerebral spinal fluid to prevent neuron degeneration. Subsequently, the brain was transversally sliced using a vibratome, and the slice was placed in an incubator for rest. After 10-15 minutes, I placed the slice in the autoclave for disinfection.
Moving on to the patch-clamp procedure, the slice was removed from the autoclave and allowed to rest in the cerebral spinal fluid for 10 minutes before being gently positioned on the brain slice rig. Microelectrodes were then utilized to puncture the membrane, and the micromanipulator was used to locate living neurons. The intracellular solution was then injected using a micropipette to prevent neuron implosion and atmospheric pressure was employed via a pump to remove debris.
When I located a living neuron, I injected it with 4-Aminopyridine. This effect blocked most of the potassium voltage-gated channels, allowing for constant depolarization for a given period of time. The first image reflects the action potentials in the absence of the drug, while the second image portrays the action potentials under its influence.
The second image clearly illustrates a period of inhibited depolarization, indicating the effect of what these inhibitory drugs and other environmental factors could cause. In the coming two weeks, I will be posting another joint post with the final details of my research paper, including its significance to future research.
See you next time!