The Practice of Patch Clamping--膜片钳操作
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1151
In this Chapter the actual procedure of making a gigaseal on a cell
membrane and establishing the desired configuration is discussed. The
purpose of this is to indicate good practice. Practitioners might find that
this Chapter helps to increase the success rate, facilitates trouble-shooting
and reduces having to ‘reinvent the wheel’. Novices hopefully will be saved
much time. The method used in this Chapter is to provide a step-by-step
account of how to establish a patch clamp configuration, using the
background information outlined in the previous Chapters. Electrophysiology
is one of the disciplines within biology that allow many instant
quality controls; these will be indicated where appropriate.
membrane and establishing the desired configuration is discussed. The
purpose of this is to indicate good practice. Practitioners might find that
this Chapter helps to increase the success rate, facilitates trouble-shooting
and reduces having to ‘reinvent the wheel’. Novices hopefully will be saved
much time. The method used in this Chapter is to provide a step-by-step
account of how to establish a patch clamp configuration, using the
background information outlined in the previous Chapters. Electrophysiology
is one of the disciplines within biology that allow many instant
quality controls; these will be indicated where appropriate.
4 The Practice of Patch Clamping 95
4.1 Preparing the Experiment and Making a Seal 95
4.1.1 Setting up 95
4.1.2 Bringing the pipette near the preparation 98
4.1.3 Making the seal 101
4.2 Whole-cell Modes 104
4.2.1 Conventional whole-cell recording 104
4.2.2 Perforated patch recording 108
4.3 Single-channel Modes 110
4.3.1 General notes 110
4.3.2 Cell-attached patch 112
4.3.3 Excised patches 113
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4.1 Preparing the Experiment and Making a Seal 95
4.1.1 Setting up 95
4.1.2 Bringing the pipette near the preparation 98
4.1.3 Making the seal 101
4.2 Whole-cell Modes 104
4.2.1 Conventional whole-cell recording 104
4.2.2 Perforated patch recording 108
4.3 Single-channel Modes 110
4.3.1 General notes 110
4.3.2 Cell-attached patch 112
4.3.3 Excised patches 113
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