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Study of bovine chromaffin cells and of their sensitivity to acetylcholine by Patch clamp (CAT#: STEM-PET-0025-WXH)

Introduction

The chromaffin cell is widely used as a model for neuroendocrine secretory cells in general and noradrenergic neurons. Indeed, the structure and function of the components of the secretory apparatus appear to be quite analogous in chromaffin cells and neurons. The chromaffin cell is a type of cell that makes neurohormones (chemicals that are made by nerve cells and used to send signals to other cells) and releases them into the blood. Chromaffin cells make epinephrine (adrenaline) and norepinephrine (noradrenaline).




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