Bad access resistance

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JennaMo
JennaMo's picture
Bad access resistance

I am attempting to patch clamp SCN (an area of the hypothalamus) cells in the whole cell configuration. I have extensive experience with HEK cells but have only been patching these cells in a brain slice for the past few months with limited sucess.

These cells are small (8-12 um) GABA-ergic neurons not organized in a laminar structure like hioppocampus and my big problem is that once I get a good seal and go whole cell the access resistance is just too high. I have tried to improve access by reapplying negative pressure or zapping but with limited success.

Does anyone have any tips or tricks for improving access to the cell? I have tried different pipettes and different techniques for going whole cell but nothing is working for me.

Thanks,

Jenna.

The FFM
The FFM's picture
 It sounds as if you are

 It sounds as if you are having issues with your patches resealing.

Have you considered using the perforated patch technique for your experiments?

see this conversation about resealing patches from a few years ago on scientist solutions for some other points

http://www.scientistsolutions.com/t2475-anyone+got+a+good+cure+to+stop+access+resistance+creeping+higher_.html

JennaMo
JennaMo's picture
Yes, I think you're right; my

Yes, I think you're right; my patches are re-sealing. No I hadn't considered perforated patch. It may be a good alternative if I just can't get the whole cell to work. Thanks.