I use regular KGluc solution, but with ajusted to 450 mOsm osmolarity. Bath solution is regular Tyrode-based, with normal osmolarity. Under these conditions gigaseal forms very quickly, but when I try to make o whole-cell configuration in regular way: by applying negative preassure ar by electric pulse usually I loose seal and cell. Does anybody knows any tips for this case?
If the problem is cell swelling due the increased intracellular osmolarity, you could try to make your bathing solution hyperosmolar, with mannitol or similar..
Thank you. But my task is to explore, how changes properties of cell during this swelling.
I think, Dr. Brodin suggests a reasonable approach: make both solution of about similar osmolality just to get giga-seal and viable WCR, and then play with osmolality of the external solution along with your experimental needs. Also, you may want to try ZAP function alone or in combination with a slight negative pressure inside pipette
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