Hello

For weeks I have issues with stability of my outside-out macro patches of oocytes expressing P2X7 receptor. In majority of cases, less than a second after ATP application patch breaks, activation is reaching steady-state and in miliseconds it breaks. Amplitudes are not high, round 200 pA, and even at 100 pA it breaks. Voltage is -60mV, 150 KCl solution EC and IC.

When the cell is empty, without receptor expression, patches are very stable for long time.

So obviously receptor is the guilty one. But what can help?

Also quite often I get patches without current from expressing cell, so I think probably vesicles are being formated inside pipette, but do not know how to fight that?