Where can I find more information on the theory and principles behind whole cell patch clamp recording? I'm recording from cultured mammalian cells transfected with recombinant receptors, and I would like to familiarize myself more with the different types of errors (e.g. voltage errors from currents that are too large), how to judge the quality of a seal based on the shape of the test pulse response, etc. as well as how the recording equipment operates (digitization, sampling rate, filtering, etc.) that applies to patch clamp recording in general.
I would strongly recommend giving the Axon Guide a read! You can download the PDF for free on the Molecular Devices website. One major voltage error to look out for is series resistance. Bill Connelly has a blog post titled "Series Resistance. Why it's bad." that explains how it affects your recordings, and a follow-up on how to properly compensate for that. To add, if you plan on compensating for series resistance, the quickest method of minimizing pipette capacitance that the lab I'm in uses is wrapping a thin strip of parafilm around the pipette until you get close to the tip and minimize the amount of internal solution. Another source of error is the liquid junction potential which is voltage offset caused by differences in cation and anion sizes in the pipette solution, which is really only notable if you use a large anion base for your solution like K-gluconate. Nat Blair has a blog titled "What is a junction potential?" and a follow-up titled "How do I measure the liquid junction potential?" that goes over it in better (and more correct) detail than I did. Sorry for not adding links, I'm not sure what CZcams's policy is for adding links on comments. I know I'm a little late on the reply, but I hope this helps! Edit: I'm not associated with this lab, but I primarily do patch for my research
Hello, where do you place your carbogen gas tank? At my university they ask me for evidence to allow me to place it next to the recording setup. Where is your lab located?
Very thorough! Including technical descriptions as well as conceptual. Thank you for this!
I can't thank you enough for such detailed information.
This is such a sophisticated experimental procedure. Thanks for this video.
This is an absolutely fantastic video thank you so much.
Great content! Thanks for this!
Excellent Content ! Very Informative !
This is great! Thank you!
Thank you for your detailed explanation.
excellent video. good job man
Thanks for this video, so interesting
great content
Nice content, Thanks.
You explained how to giga seal in details and perfectly,
Very nice Thank you
Great video and very informative! Is the rest of the video online too?
What book teaches how to read voltage clamp recordings ?!
Where can I find more information on the theory and principles behind whole cell patch clamp recording? I'm recording from cultured mammalian cells transfected with recombinant receptors, and I would like to familiarize myself more with the different types of errors (e.g. voltage errors from currents that are too large), how to judge the quality of a seal based on the shape of the test pulse response, etc. as well as how the recording equipment operates (digitization, sampling rate, filtering, etc.) that applies to patch clamp recording in general.
I would strongly recommend giving the Axon Guide a read! You can download the PDF for free on the Molecular Devices website.
One major voltage error to look out for is series resistance. Bill Connelly has a blog post titled "Series Resistance. Why it's bad." that explains how it affects your recordings, and a follow-up on how to properly compensate for that. To add, if you plan on compensating for series resistance, the quickest method of minimizing pipette capacitance that the lab I'm in uses is wrapping a thin strip of parafilm around the pipette until you get close to the tip and minimize the amount of internal solution.
Another source of error is the liquid junction potential which is voltage offset caused by differences in cation and anion sizes in the pipette solution, which is really only notable if you use a large anion base for your solution like K-gluconate. Nat Blair has a blog titled "What is a junction potential?" and a follow-up titled "How do I measure the liquid junction potential?" that goes over it in better (and more correct) detail than I did.
Sorry for not adding links, I'm not sure what CZcams's policy is for adding links on comments. I know I'm a little late on the reply, but I hope this helps!
Edit: I'm not associated with this lab, but I primarily do patch for my research
muito pika
Hello, where do you place your carbogen gas tank? At my university they ask me for evidence to allow me to place it next to the recording setup. Where is your lab located?
We have portable attachments that allow us to connect it to the bench top securely.