Electron microscopic radioautography of thin sections: the Golgi zone as a site of protein concentration in pancreatic acinar cells.

Abstract

Electron microscopic radioautographs of guinea pig pancreatic exocrine cells were obtained by covering thin sections ( approximately 600 A) of OsO(4)-fixed, methacrylate-embedded tissue with thin layers of Ilford K-5 nuclear research emulsion. After an exposure of 13 days at 4 degrees C., the preparations were photographically processed, stained with uranyl acetate, and examined in an electron microscope. The label used was leucine-H(3) injected intravenously 20 minutes before collection of the specimens. Conventional radioautographs of thicker sections (0.4 micron) were also examined in a phase contrast microscope. The advantages obtained from electron microscopic radioautography are: the higher radioautographic resolution (of the order of 0.3 micron) due to the thinness of the emulsion and the specimen, and a high optical resolution permitting a clear identification of the labeled structure. In the guinea pig pancreas this technique demonstrated that, at the time studied, newly synthesized proteins were concentrated in the structures of the Golgi complex and especially in large vacuoles partially filled with a dense material. The vacuoles are probably a precursor to the secretion granules (zymogen granules) in which the label becomes segregated at a later time. These observations demonstrate directly the role of the Golgi complex in the secretion process. They also illustrate the possibilities of this method for radioautography at the intracellular level.