Patch Clamp Electrophysiology

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.

Patch-clamp workflow using Axon Instruments

The Axon Instruments® portfolio provides comprehensive solutions for patch-clamping that includes amplifiers, digitizer, software, and accessories. Our best-in-class instruments facilitate the entire range of patch-clamp technique experiments from the smallest single channel to the largest macroscopic recordings. The addition of Axon pCLAMP 11 Software Suite creates a streamlined workflow, allowing for sophisticated and effecient experiments, and higher quality data generation Learn more about the electrophysiology lab set up >

Patch-clamp workflow using Axon Instruments

  1. Prepare solutions – Make internal and external solutions. Adjust osmolarity and pH values.
  2. Prepare cells or brain slices – Prepare cultured cells, isolated neurons, brain slices, or whole animals.
  3. Pull and polish the pipette – Prepare the recording electrode. Pull the glass capillary tube and polish pipette tip.
  4. Set up the perfusion system – Set up the perfusion system and data acquisition software. Ensure that the system is shielded.
  5. Patching a cell – Use the manipulator to touch the cell membrane with the pipette. Ensure a high resistance electrical seal is formed.
  6. Signal acquisition and amplification – The signal will be amplified. For best results, ensure you are using the correct type of amplifier for your research.
  7. Signal digitization – The analog signal is then digitized so that the signal can be analyzed.
  8. Data acquisition and analysis – With pCLAMP 11 Software Suite, longer and more sophisticated protocols can be programmed for faster data analysis and precise measurements.

Learn about electrophysiology lab set-up

Learn more about patch clamping techniques, from single channel to whole cell to extracellular field-potential recording.

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