prevent cell reseal after going whoole cell

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Sultan S. ALSiyabi
Sultan S. ALSiyabi's picture
prevent cell reseal after going whoole cell

Hi, I would like you to advice me how to prevent cell reseal after going whoole cell?
I am working with SMCs isolated from coronary artery.

Fraser Moss
Fraser Moss's picture
The best way I have found to

The best way I have found to avoid resealing is to spend time designing a new program on the puller that pulls wider tips that still have the desired resistance when filled with intracellular solution. How wide are you tips? See if you make them at least 1.5 microns inner diameter at the tip.

Another trick if you have not already done so is to increase the EGTA concentration to 10mM in your pipette solution.