The Patch Guide
A Student's Guide to Patch-Clamp Electrophysiology

An introduction to basic concepts required to perform and evaluate whole-cell patch clamp experiments
  • Getting Started - How to use this guide
  • Introduction - Thinking about neurons as electrically excitable membranes
  • Passive Properties - Electrical features of a cell membrane that do not require channels to actively open or close
  • Ohm’s Law - How Membrane Properties Influence Synaptic Currents
  • Action Potentials - Electrophysiology of neuron firing
  • VC and IC Modes - Voltage-clamp and current-clamp whole-cell recording configurations
  • Synaptic Currents - Electrical changes in a cell resulting from neurotransmitter released by another cell
  • How to Patch Clamp Neurons - Steps for establishing whole-cell patch-clamp configuration with neurons in brain slices
Individual pages for a variety of topics
  • Common Experiments - A summary of the most common whole-cell patch-clamp experiment types
  • Common Pharmacological Agents - Pharmacological agents commonly used in whole-cell patch-clamp experiments
  • Electrically Evoke Synapses - Using an electrical stimulator to evoke synaptic currents
  • Liquid Junction Potential - What Liquid Junction Potential (LJP) is and how it can be corrected for in patch-clamp experiments
  • Optogenetics - Using light-activated ion channels to optically excite specific cell populations
  • Patch Clamp Internal Solutions - Common internal solutions used for whole-cell patch-clamp electrophysiology
  • Patch Configurations - A summary of the most common techniques used to measure the electrical properties of cell membranes
  • Reversal Potential - An introduction to reversal potentials, the Nernst equation, and their effect on synaptic currents
  • Shunting - Synaptic inputs can be attenuated in a process known as current shunting
  • The Membrane Test - A voltage-clamp step can reveal much about a neuron's passive membrane properties
Resources for analyzing electrophysiological and optical data
Personal notes about hardware and software which may be laboratory-specific
  • 2P Operation Notes - Notes about two-photon startup, shutdown, acquisition, and maintenance
  • 2P Stage Control - Python scripts to automate two-photon stage positioning, image acquisition, and montage assembly using ImageJ and TrackEM
  • Aligning the Mira Laser - Notes related to power and alignment of a Coherent Mira Optima 900-F
  • DIC Alignment Cheat-Sheet - How to align DIC optics for a fixed-stage Olympus BX-51WI microscope
  • NT-RTSI Setup - National Instruments Real-Time System Integration System Setup
  • Pipette Puller - How to make whole-cell patch-clamp pipettes using the P97 Flaming/Brown pipette puller
  • Preamp Blade Setup - How to configure and switch between different preamplifier blades
  • Two-Photon Laser Uncaging - How to setup, calibrate, and run mixed ephys/imaging/uncaging experiments
  • Two-Photon Linescans - How to acquire fast imaging data using two-photon linescans