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Drug discovery by Automated patch clamp by Patch clamp (CAT#: STEM-PET-0030-WXH)

Introduction

Patch-clamp electrophysiology remains an important technique in studying ion channels; indeed, it is still considered the gold standard since it was first described by Neher and Sakmann in the 1970s. Ion channels are integral membrane proteins which allow ion current flow across the cell membrane. They are involved in almost all physiological processes, and their malfunction underlies many disease states, making them important pharmacological targets.




Principle

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.

Applications

• Study of ionic currents in individual isolated living cells, tissue sections, or patches of cell membrane.
• Study of excitable cells such as neurons, cardiomyocytes, muscle fibers, and pancreatic beta cells.
• Study of ion channels.

Procedure

1. Fabrication of glass electrodes
2. Measuring glass electrode resistance and compensating offset potential
3. Glass electrode contact to cell membrane and obtain a GΩ seal
4. Acquire and analyse recordings using the appropriate software.

Materials

Patch clamp system