Loosing seal going whole cell

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frogger
frogger's picture
Loosing seal going whole cell

Hi there,
I have been trying to patch from Purkinje neurons and am having difficulty. I can seal the cells very consistently, however when I apply suction, the seal drops immediately down to less than 100 MOs nearly every time. I am not sure if it is the quality of the cells or a problem with the solutions.  The internal solution has an osmolarity of 290, and the external (Tyrode's) is at 320. Any suggestions would be immensely appreciated!
Thanks so much!

lazy
lazy's picture
When you apply suction, do

When you apply suction, do you keep the electrode potential near the restig potential? (I am assuming that you use high K internal solution. )
Or, do you use gluconate (or something else instead of K)  internal solution? In such case, I doubt the cell.