Good Day.
I am doing patch-clamp electrophysiology of ligand-gated ion channels (LGIC) in HEK293 cells, and I am still working my way to understand the technical details of managing the perfusion system.
Recently, I observed that we have a slow dropping of the solution in the perfusion manifold at 2psi. This is reflected in my recordings of saturating agonist concentration. This is because I did not observe a fast onset of the LGIC. Instead, it has slow activation which is not similar based on the current peaks shapes of the published literature at saturating GABA concentration.
We think that there might be a bubble or clogging along the lines of the perfusion system. Right now, we are doing ejection of the perfusion lines in at high pressure system to ensure that we remove the bubble or any clogging.
In your case, can you suggest some other methods to remove clogging/bubble so we could minimize or eliminate its effect when I record cells next time?
I would also like to ask if there is a manual or paper about managing the perfusion system in electrophysiology? This would help me understand more about the technical details of the ephys setup aside from the company's technical manual.
Hoping for your insightful answers.
Thank you very much.
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