16 June 2021 3 721 Report

Hi everyone,

I'm new at doing perforated patching, and I am having a difficult time obtaining the gigaseal.

Here are some information and questions that I have:

  • The concentration of amphotericin B I use is 300 μg ml−1
  • I'm using dissociated neurons and trying to do a current clamp.
  • I try to fill the tip with an amphotericin B-free internal solution, but I'm having difficulty for some reason. the pipette resistance is between 2.5 to 4 MΩ (so maybe the tip is too small to fill with a tube & syringe?)
  • Previously, I used to do whole cell patching, and we measured the osmolarity of our internal and external solution (where our internal solution is 10 mOsm less than the external solution). But the person I'm learning perforated patch from says it does not matter. Plus, we don't have an osmometer to measure them. So does internal and external solution only matter when I do whole cell patching, not perforated patching?
  • Thank you in advance, and any advice is appreciated

    Joyce

    More Joyce Kim's questions See All
    Similar questions and discussions