Patch Clamp Electrophysiology

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Patch clamp electrophysiology

The Patch-clamp technique is a versatile electrophysiological tool for understanding ion channel behavior. Every cell expresses ion channels, but the most common cells to study with patch-clamp techniques include neurons, muscle fibers, cardiomyocytes, and oocytes overexpressing single ion channels.

To evaluate single ion channel conductance, a microelectrode forms a high resistance seal with the cellular membrane, and a patch of cell membrane containing the ion channel of interest is removed. Alternatively, while the microelectrode is sealed to the cell membrane, this small patch can be ruptured giving the electrode electrical access to the whole cell. Voltage is then applied, forming a voltage clamp, and membrane current is measured. Current clamp can also be used to measure changes in membrane voltage called membrane potential. Voltage or current change within cell membranes can be altered by applying compounds to block or open channels. These techniques enable researchers to understand how ion channels behave both in normal and disease states and how different drugs, ions, or other analytes can modify these conditions.

Quick links to Patch clamp electrophysiology basics:

  • Action Potential

    what-is-action-potential

     An action potential is a rapid rise and subsequent fall in voltage or membrane potential across a cellular membrane with a characteristic pattern. Examples of cells that signal via action potentials are neurons and muscle cells.

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    Cellular Pathway Analysis

     Cellular Pathway Analysis

    Ion channels are involved in many cell pathways and understanding the function of ion channels in response to changes in membrane potential or the presence or absence of other molecules is important in order to understand exactly how ion channels participate in normal and abnormal biological processes such as cell differentiation and migration, disease states, and neuronal communications.

  • cSEVC

    What is continuous single electrode voltage-clamp (cSEVC)?

    What is continuous single-electrode voltage clamp (cSEVC)? It is an electrophysiology patch-clamp method that passes a membrane voltage into a cell and measures the change in current as the voltage steps.

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    Current Clamp Amplifier

    Current Clamp Amplifier

    Current-clamp is a method used to measure the resulting membrane potential (voltage) from an injection of current. To measure the membrane potential, the MultiClamp 700B and Axoclamp 900A both monitor voltage drop initiated by current injection along an in-series resistor. Current-clamp is commonly used to inject simulated, but realistic current waveforms into a cell, and monitor membrane effect. This technique is ideal for the evaluation of important cellular events such as action potentials.

  • Digital Acquisition

    Patch-Clamp Electrophysiology Digital Acquisition

    The current or voltage signal acquired by the amplifier is an analog signal, but to perform data analysis needed for high resolution patch-clamp measurements, the analog signal must be converted into a digital one. Positioned between the amplifier and the computer, the digitizer accomplishes this important task. Signal quality is extremely important and is impacted by the sampling frequency. The latest generation of Digidata digitizers sample at 500 kHz and can be equipped with HumSilencer, which eliminates 50/60 Hz line-frequency noise.

    Disease Research

    Ion Channels used in Disease Research

    Ion channels play a role in many diseases including hypertension, cardiac arrhythmias, gastrointestinal, immune and neuromuscular disorders, pathological pain, and cancer. By understanding the exact role that ion channel play in a particular disease, researchers might be able to find a way to affect the ion channel in such a way as to alter the course of the disease.

  • dSEVC

    What is discontinuous single electrode voltage-clamp (dSEVC)?

     In discontinuous single-electrode voltage clamp (dSEVC), the tasks of voltage recording and current passing are allocated to the same micropipette.

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    Electrophysiology

    Electrophysiology: Patch Clamp

    Electrophysiology is one of the foundational disciplines in neuroscience and cardiac physiology for the evaluation of ion channels. The patch-clamp technique is a versatile electrophysiological tool for understanding ion channel behavior.

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  • Ion Channels

    Ion Channels

    An ion channel is a group of proteins that form a pore across the lipid bilayer of a cell. Each channel is permeable to a specific ion (examples: potassium, sodium, calcium, chloride). Patch-clamp is used to evaluate current or voltage in the membrane associated with ion channel activity via direct measurement in real time using ultra-sensitive amplifiers, high-quality data acquisition systems, and powerful software to evaluate the results.

    Patch Clamp

    Patch-Clamp Electrophysiology Techniques

    The patch-clamp technique involves a glass micropipette forming a tight gigaohm (GΩ) seal with the cell membrane. The micropipette contains a wire bathed in an electrolytic solution to conduct ions. The whole-cell technique involves rupturing a patch of membrane with mild suction to provide low-resistance electrical access, allowing control of transmembrane voltage. Alternatively, investigators can pull a patch of membrane away from the cell and evaluate currents through single channels via the inside-out or outside-out patch-clamp technique.

  • Series Resistance Compensation

    Series Resistance Compensation using Whole-Cell Recording Method

    Series resistance is the sum of all resistances between the amplifier and the inside of the cell using the whole-cell recording method. Due to Ohms Law, the larger this resistance, the greater the difference between the command level and the measured values. This creates an error in actual voltage or current measurement potentially leading to inaccurate observations. To overcome this, the Molecular Devices amplifiers have built-in circuitry to improve the bandwidth of the recording by compensating the error introduced by the voltage or current drop across the series resistance.

    Single Channel Recording

    Patch-Clamp Single Channel Recording Technique

    The patch-clamp technique involves a glass micropipette forming a tight gigaohm seal with the cell membrane. The micropipette contains a wire bathed in an electrolytic solution to conduct ions. To measure single ion channels, a “patch” of membrane is pulled away from the cell after forming a gigaohm seal. If a single ion channel is within the patch, currents can be measured. The Axopatch 200B, with extremely low-noise profile, is ideal for this application, maximizing signal for the smallest conductance ion channels.

  • The Axon Guide

    The Axon Guide

    A guide to Electrophysiology and Biophysics Laboratory Techniques. The purpose of this guide is to serve as an information and data resource for electrophysiologists. It covers a broad scope of topics ranging from the biological basis of bioelectricity and a description of the basic experimental setup to a discussion of mechanisms of noise and data analysis.

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    Voltage Clamp Amplifier

    Voltage Clamp Amplifier

    In an experiment using the voltage-clamp method, the investigator controls the membrane voltage in a cell and measures the transmembrane current required to maintain that voltage. This voltage control is called a command voltage. To maintain this command voltage level, an amplifier must inject current. The current injected will be equal and opposite the current escaping through open ion channels, allowing the amplifier to measure the amount of current passing through open membrane bound ion channels.

  • Whole Cell Recording

    Whole Cell Recording Patch-Clamp Technique

    The whole cell patch-clamp technique involves a glass micropipette forming a tight gigaohm (GΩ) seal with the cell membrane. This micropipette contains a wire bathed in an electrolytic solution to conduct ions. A patch of membrane is subsequently ruptured by mild suction so that the glass micropipette provides a low-resistance access to the whole cell, thereby allowing the investigator to control the transmembrane voltage and allowing the investigator to evaluate the sum of all currents through membrane bound ion channels.

Resources of Electrophysiology

Videos and Demos

Calculate Decay Time Constant, and Perform Curve Fitting Using Axon pCLAMP Software

How to Combine Traces, Calculate Rise or Decay Time Constant, and Perform Curve Fitting Using Axon pCLAMP Software

Create Customized Command Waveforms Using pCLAMP Software

How to Create Customized Command Waveforms Using the pCLAMP Software

Use of Sequencing Keys, User List, and Stimulus File with pCLAMP Software

The Use of Sequencing Keys, User List, and Stimulus File with pCLAMP Software

Synchronizing Electrophysiology and Imaging Solutions with pCLAMP and MetaMorph Software

Synchronizing Electrophysiology and Imaging Solution with Axon pCLAMP and MetaMorph Software

Membrane Test Between Sweeps in Clampex & Analysis of Synaptic Events with Clampfit Data Analysis

Online Statistics, Membrane Test Between Sweeps in Clampex and Analysis of Synaptic Events with the Clampfit™ Data Analysis

Use of Axoporator for Single-cell Electroporation for Transfection and Dye-labeling

Use of the Axoporator 800A for Single-cell Electroporation for Transfection and Dye-labeling

Using Axoclamp 900A for Two-Electrode Voltage-Clamp of Xenopus Oocytes Expressing Ion Channels

Using the Axoclamp 900A for Two-Electrode Voltage-Clamp of Xenopus Oocytes Expressing Ion Channels

Use of Filters in Data Acquisition and the Clampfit Application

Writing Long-Term Potentiation and Depression Protocols and the Use of Filters in Data Acquisition and the Clampfit Application

Resistance Compensated Series

Series Resistance Compensated or Not

Using Electrophysiological Studies to Accelerate Mechanistic in Reception and Transmission

Using Electrophysiological Studies to Accelerate Mechanistic Study in Reception and Transmission

Hardware Choices for Optogenetics Considerations for Synchronized Light Patterning

Update and Hardware Choices for Optogenetics Considerations for Synchronized Light Patterning

Effects of Amyloid-Beta Proteins on hSlo1.1, a BK Channel, in Xenopus Oocyte Model

Investigations of the Effects of Amyloid-Beta Proteins on hSlo1.1, a BK Channel, in a Xenopus Oocyte Model

Nanopores-Electronic Tools for Single-Molecule Biophysics and Bio-Nanotechnologies

Nanopores-Electronic Tools for Single-Molecule Biophysics and Bio-Nanotechnologies

Axon Amplifiers and pCLAMP Software-key Features Reviews (Chinese version)

Axon Amplifiers and pCLAMP Software-key Features Reviews (Chinese version)

Using Clampfit in Basic Single Channel Analysis

Basic Single Channel Analysis Using Clampfit

Action Potential Analysis in Clampfit Module

Action Potential Analysis in Clampfit Module

A Walkthrough of Protocol Editor in pCLAMP Data Acquisition Module

A Walkthrough of the Protocol Editor in the pCLAMP Data Acquisition Module

A Walkthrough of Protocol Editor in pCLAMP (Chinese version)

A Walkthrough of Protocol Editor in pCLAMP (Chinese version)

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