心脏电生理技术基础(膜片钳).ppt

CELLULAR CARDIAC ELECTROPHYSIOLOGICAL TECHNIQUES NORBERT JOST, PhD Patch-clamp: the special case of the voltage clamp Patch-clamp: the special case of the voltage clamp Patch-clamp: the special case of the voltage clamp Patch-clamp: the special case of the voltage clamp Patch-clamp: the special case of the voltage clamp Properties of individual voltage-dependent sodium channels Properties of individual voltage-dependent sodium channels Properties of individual voltage-dependent sodium channels Properties of individual voltage-dependent sodium channels Properties of individual voltage-dependent sodium channels Properties of individual voltage-dependent sodium channels Similarly, individual potassium channels, calcium channels, and other channels can be studied by patch clamping The configurations of the patch-clamp technique The configurations of the patch-clamp technique The configurations of the patch-clamp technique The configurations of the patch-clamp technique The configurations of the patch-clamp technique The configurations of the patch clamp technique Whole Cell Whole Cell outside-out patch Rs Cm Rc The whole-cell configuration NaCl 144 NaH2PO4 0.4 KCl 4 MgSO4 0.53 CaCl2 1.8 Glucose 5.5 HEPES 5 + ICa blocker Intracellukar solution (mM) (for K currents) Extracellular solution (mM) (for K currents) K-aspartate 100 KCl 25 K2HPO4 10, K2EGTA 5 K2ATP 3 MgCl2 1 HEPES 10 Extracellular solution Patch-clamp amplifier IBM PC Micropipette + _ _ + + + + + _ _ _ _ _ + + _ _ + + _ Cell -40 mV -20 mV ... +50 mV 10 ms ... 5000 ms Intracellular solution The whole cell configuration The “run-down“ effect The ATP-sensitive potassium current The L-type calcium current The “run-down“ Whole Cell Whole Cell, perforated patch - amphotericin-B - nystatin * Electrical model of the membrane Standard intracellular microelectrode technique Voltage clamp technique Patch clamp technique G=1/R Ohm’s law Ion channel model Current clamp