Hi all,
I'm curious to see if anyone has experience using Sylgard or Parafilm to coat their pipettes when doing whole-cell voltage clamp. I found that this technique has frequently been present since the late 90s on voltage clamp papers, and I'm wondering if it's worth doing to reduce capacitance on my own cells. I am patching on Purkinje neurons in both dissociation and slice preparations, and recording INa current.
If someone could point me to a paper or just describe their experiences here it would be greatly appreciated! I'm not exactly sure where to start (how much to add? where to add? what techniques work best? do you apply either under a dissection scope?) so please be descriptive as possible.
Thank you!
I cover the microelectrode with a hot glue gun (see image). I simply rotate the patch pipette wall in the melted glue and wait for it to cool (a few seconds). It takes some practice to cover the patch pipette nicely. This plastic glue significantly reduces the stray capacitance caused by the patch pipette glass.
Hope this helps,
Albertino
Dear Samuel Brown ,
In my experience, I've seen people use sylgard, dental wax and parafilm (and now, hot glue with Albertino Bigiani !). From my own experience:
- Sylgard needs to be mixed and cured, so you need to wait for it or heat it to cure faster;
- Wax requires some practice to make a good coating and you need to use it while hot - it is solid at room temp;
- Parafilm works as a charm and I think it's the easiest.
I've used parafilm already and I recommend as a starting point due to the convenience of it. There's a post from Lina Kenzler (How to get Sylgard right for coating ? - https://www.researchgate.net/post/How_to_get_Sylgard_right_for_coating ) where I put pictures of it.
Please feel free to reach out if you need some more clarifications.
Good luck!
Andre
ps: I used wax too, but I did not have the skill for it, so the frustration drove me to the parafilm!
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