I have been trying to patch transfected cos-7 cells (Kv channel plasmid) and am having an incredibly difficult time getting a seal let alone breaking in. Has anyone every patched with these cells and seen similar issues? Or in regards to transfection reagents (currently using LTX). I really don't think it's my extracellular or intracellular because I am using previously published protocols.
COS-7 cells shouldn't be a problem for patch clamping, most cultured cells are friendly. Did you succeed with other cells? untransfected COS cells? Any special reason for using them and not HEK293 cells?
I agree with Maximilian, cells from cell lines are friendly, but it can be tricky to find the best method to get a gigaseal. I am working with cells from a bronchial cell line, and it took some time before I could get any gigaseal. It was primarily myself :) I wasn´t doing things properly or consistently. The most critical part is to put the tip of the pipette in a good place on the middle of the cell so that suction will have enough membrane to "drag into" the tip (or something of that sort). Sometimes it seems suction isn´t doing anything, and you wonder if something is wrong with the holder or the line. Now for instance I am not getting consistent gigaseals, but it is because I lost on practice during the holidays :)
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