A "skinning%" procedure is described for irreversibly disrupting the sarcolemmal membrane of human skeletal muscle and allowing calcium and other diffusible solutes (such as adenosine triphosphate) access to the myofilament space. Single skinned fibers give isometric tensions of about 1.5 kilograms per square centimeter when exposed to ionized calcium event after 1 to 2 weeks of storage at 5 degrees C. For up to 5 days the preparation will sequester and, under appropriate conditions (anion substitution, caffeine addition, or magnesium withdrawal), release calciumn. The regulation of intracellular calcium distribution and the calcium-induced activation of the contractile proteins are discussed and related to the morphology of humnan fibers and to similar processes occurring on other muscle preparations.