If anybody could give me advice I would really appreciate it.
I am trying to patch pyramidal and granule cells in acute hippocampal slices. I get about 3-4GOhm seal quickly but it does not hold. It breaks away in <1min or so. If I apply -70mV, as suggested to make better seal, it breaks away even faster. In cultures I did not have this problem, it seals there up to 15GOhm and stays fro a long time. My bath solution is normal ACSF, pipette solution, 110 KGluconate, 10 KCl, 2mgMgATP, 10 EGTA, 10HEPES pH 7.2, Osm 290mOsm, R=6MOhm.
Thank you in advance
acute hippocampal slices