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J Vis Exp. 2008 Oct 16;(20). pii: 936. doi: 10.3791/936.

Patch clamp recording of ion channels expressed in Xenopus oocytes.

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1
Department of Molecular and Cellular Physiology, Stanford University, Stanford, CA, USA. austin.brown@stanford.edu

Abstract

Since its development by Sakmann and Neher (1, 2), the patch clamp has become established as an extremely useful technique for electrophysiological measurement of single or multiple ion channels in cells. This technique can be applied to ion channels in both their native environment and expressed in heterologous cells, such as oocytes harvested from the African clawed frog, Xenopus laevis. Here, we describe the well-established technique of patch clamp recording from Xenopus oocytes. This technique is used to measure the properties of expressed ion channels either in populations (macropatch) or individually (single-channel recording). We focus on techniques to maximize the quality of oocyte preparation and seal generation. With all factors optimized, this technique gives a probability of successful seal generation over 90 percent. The process may be optimized differently by every researcher based on the factors he or she finds most important, and we present the approach that have lead to the greatest success in our hands.

PMID:
19078941
PMCID:
PMC3234036
DOI:
10.3791/936
[Indexed for MEDLINE]
Free PMC Article
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