Loss of gigaseal

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Nasiara's picture
Loss of gigaseal

I need some help and I really dont have amyone around to offer that. I am working on cultures hippocampal neurons in inside out patches recording. I am new to the techinique. I regularly get gigaseals but by the time I switch the gain to 500 (with holding potential -60mv) on Axopatch 200A, the seal is lost.
Even if it does not on that point, after half a minute or a minute the seal breaks. I was just wondering if you could give some advice why it so? Thanks