simple questions about patch clamp

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keyong's picture
simple questions about patch clamp

Two simple questions are confusing me.
1) In membrane test window, should the baseline of step pulse be set at zero or the middle of step pulse at zero when the pipette is moved into solution?

2) If a negative holding potential is set in order to easily get Giga seal, does it affect the membrane potential when switched to current clamp?
Thanks a lot!

stovepower's picture
 Hi climber,

 Hi climber,

Baseline of the step pulse should be set to zero.

It should not affect membrane potential when switched back to current clamp if your holding potential is pretty close to membrane potential already. In our lab we use a holding potential of -80 mV, which works fine even though the membrane potential for our neurons may be closer to -90 mV.