X-ray microanalysis of cryofixed skeletal muscle in culture

Abstract

X-ray microanalysis of electrolyte ions in cells and tissues involves the problem of preserving the distribution of diffusible substances throughout preparation and investigation in the scanning electron microscope. Cryofixation of objects thicker than about 200,um induces considerable freezing artifacts. The dissection of the specimen from the intact tissue also influences the distribution of diffusible ions. With respect to these problems, tissue culture cells seem advantageous, because they need not be dissected from their environment and are small enough to be frozen without ice crystal damage.Tissue cultures were grown from skeletal muscle of the hindleg of 20-day-old rat embryos on plastic coverslips. Small pieces of the specimens (diameter about 4 mm) were mounted on cupreous specimen stubs with conducting silver paint and frozen in liquid propane.</jats:p