Patch Clamping (PDF)

Preface ix 1 Introduction 1 1.1 Patch Clamping and its Context 1 2 Basic Theoretical Principles 5 2.1 Introduction to Membrane Biology 5 2.1.1 The plasma membrane and its ionic environment 5 2.1.2 Electrochemical gradients and the Nernst equation 7 2.1.3 Maintenance of ion gradients and the membrane potential 8 2.1.4 Ion channels 11 2.2 Electrical Properties of the Cell Membrane 13 2.2.1 Driving force and membrane resistance 13 2.2.2 Membrane capacitance 15 2.2.3 Consequences of membrane capacitance 16 2.2.4 An electronic model of the plasma membrane 17 2.3 Recording Modes and their Equivalent Circuits 18 2.3.1 The basics of equivalent circuits 18 2.3.2 Intracellular recording 2.3.3 Voltage clamp and current clamp 28 2.3.4 Introduction to patch clamp configurations 2.3.5 The equivalent circuit for the cell-attached patch configuration 2.3.6 The equivalent circuit for the whole-cell configuration 39 2.3.7 The equivalent circuit for the excised patch configurations 3 Requirements 43 3.1 The Platform 43 3.1.1 Stability: vibrations and drift 43 3.1.2 Where in the building should the set-up be placed? 44 3.1.3 Anti-vibration tables 45 3.2 Mechanics and Optics 47 3.2.1 The microscope 48 3.2.2 Micromanipulators 52 3.2.3 Pipette pressure 56 3.2.4 Baths and superfusion systems 57 3.3 Electrodes and Micropipettes 64 3.3.1 Solid-liquid junction potentials and polarisation 65 3.3.2 The bath electrode 67 3.3.3 Micropipettes 67 3.3.4 Liquid junction potentials 74 3.4 Electronics 75 3.4.1 External noise and Faraday cages 76 3.4.2 Patch clamp amplifiers 81 3.4.3 Noise prevention and signal conditioning 84 3.4.4 Data acquisition and digitisation 90 3.4.5 Computers and software 93 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

Areles Molleman is the author of Patch Clamping: An Introductory Guide to Patch Clamp Electrophysiology, published by Wiley.

Dieses eBook können Sie uneingeschränkt auf allen Geräten der tolino Familie lesen. Zum Lesen auf sonstigen eReadern und am PC benötigen Sie eine Adobe ID.