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MATERIALS AND METHODS THE VOLTAGE CLAMP METHOD facilitates an understandg of the basis of dynamic electrical phenomena of the membrane. This method has heretofore not been successfully applied to muscle tissue, because of the two- or three-dimensional electrical spread of the current, large extracellular space caused by multicellular tissues, and a strg-like cell shape with an extremely small diameter (10). To overcome these difficulties, we applied the voltage clamp method for studyg dispersed sgle cell preparations. 0363-6143/86 $1.50 Copyright Preparation of dispersed sgle cells. Albo rabbits (1.8-2.2 kg) were anesthetized with pentobarbital sodium (40 mg/kg iv) and exsanguated. The longitudal muscle layers were manually peeled from the ileum. Dispersion procedures of the muscle tissue were similar to those reported (19). Briefly, small segments of the tissue were cubated for 20 m Ca2+-free physiological salt solution (PSS) at 36 t 1°C; the tissue was then soaked cubation solution contag 0.1% collagenase, 0.1% tryps hibitor, and 0.2% bove serum album. After the completion of digestion, sgle cells were dispersed by gentle agitation with a glass pipette. Debris was removed by filtration through a fe nylon mesh, and the cell suspension was centrifuged at 1,000 rpm for 1 m. The cell
AJP - Cell Physiology – The American Physiological Society
Published: Sep 1, 1986
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