Single-Cell Neuronal Dissociation for Electrophysiological Studies
- 3.9k Downloads
Since the patch-clamp technique was initially applied about 35 years ago, it has been thought to be a powerful method for studying electrical properties of ion channels and cell excitability in the varied excitable cells including peripheral and central neurons. Although we understand that many factors can affect the patch-clamp recording, the viability of the peripheral and central neurons is an essential point for the success of the patch-clamp experiments. In this chapter, I introduce (1) acute dissociation procedures of the peripheral neurons from ganglia and the central neurons from brain (such as equipment, materials, preparation of tissues, enzymatic treatment of the tissues, mechanical agitation of the tissue fragments, and preservation of the isolated cells), (2) explanatory notes for the whole procedure of the acute neuronal isolation, and (3) advantages and disadvantages of the isolated peripheral and central neurons for the electrophysiological studies.
KeywordsAction potential Cell culture Cell isolation Central nervous system Electrophysiology Ion channel Patch clamp Peripheral nervous system
- Aras MA, Hartnett KA, Aizenman E (2008) Assessment of cell viability in primary neuronal cultures. Curr Protoc Neurosci Chapter 7:Unit 7.18.1–7.18.15Google Scholar
- Kay AR, Krupa DJ (2001) Acute isolation of neurons from the mature mammalian central nervous system. Curr Protoc Neurosci Chapter 6:Unit 6.5.1–6.5.7Google Scholar