Description

This image is from a tomogram through part of a Golgi ribbon from a normal rat kidney cell, generated using high voltage electron microscopy following incubation at 15°C to block transport out of the Golgi complex. Following temperature block, large bulging domains appear on the three trans-most cisternae. The sample was tilted at angles of 1.5° to obtain the series of images in this set, and used to obtain 3-dimensional reconstructions of the Golgi apparatus and contributed to findings reported in Ladinsky et al. (1999) Golgi structure in three dimensions: Functional insights from the normal rat kidney cell. J. Cell Biol. 144 (6) 1135-1149.

Technical Details

Cells were grown on 100-mesh gold EM grids, and maintained at 15°C for 4 hours before plunge freezing in liquid nitrogen (BalTec HPM-010), followed by freeze-substitution (1% glutaraldehyde, 0.1% tannic acid in acetone, replaced with 4% osmium tetroxide and 0.01% uranyl acetate), then embedded in Epon-Araldite and sectioned at 250nm (UltraCut-UCT, Leica). Sections were transferred to formvar-coated copper-rhodium slot grids (EMS) and stained with 2% aqueous uranyl acetate and Reynold's lead citrate. Digital images were acquired at a magnification of 15,000X with an FEI Tecnai TF20 transmission EM. For additional details refer to: J. Cell Biol. 144 (6) 1135-1149.