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Whole-cell voltage clamp on skeletal muscle fibers with the silicone-clamp technique
Romain Lefebvre1, Sandrine Pouvreau, Claude Collet
1Centre de Génétique et de Physiologie Moléculaire et Cellulaire, UMR CNRS 5534, Université Claude Bernard - Lyon 1, Bât. Raphael Dubois, 43 boulevard du 11 novembre 1918, Villeurbanne Cedex, 69622, France.
Abstract:
Control of membrane voltage and membrane current measurements are of critical importance for the study of numerous aspects of skeletal muscle physiology and pathophysiology. The silicone-clamp technique makes use of a conventional patch-clamp apparatus to achieve whole-cell voltage clamp of a restricted portion of a fully differentiated adult skeletal muscle fiber. The major part of an isolated muscle fiber is insulated from the extracellular medium with silicone grease and the tip of a single microelectrode connected to the amplifier is then inserted within the fiber through the silicone layer. The method is extremely easy to implement. It represents an alternative to the traditional vaseline-gap isolation and two or three microelectrodes voltage-clamp techniques. The present chapter reviews the benefits of the silicone-clamp technique and provides updated detailed insights into its practical implementation.
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