I am having a lot of difficulty lately patching acutely isolated neurons from CA1 region of hippocampus. I have no trouble with sealing and breaking through, the problem arises when I lift the cell (after breakthrough) off the plate. Shortly after lifting the cell into the stream of solution, the cell falls off the electrode. I wonder if the problem has to do with the seal itself, the osmolarity of my solutions, CsF in my ICF....
The ECF of my solutions is usually 325-300 mOsm. The ICF is usually between 295-300 mOsm.
Please let me know if you have any suggestions.