CIL:13760, Rattus norvegicus, pheochromocytoma, adrenal gland cell. In Cell Image Library

Electron micrograph of PC12 cells transfected with control siRNA. PC12 cells were plated onto aclar film discs (Pella) coated with poly-L-lysine, transfected twice with 50 nM control siRNA and fixed with 2.5% glutaraldehyde in calcium-/magnesium-free PBS. The discs were washed three times with 0.1 M sodium cacodylate buffer, pH 7.2, and postfixed for 30 min on ice with 1% osmium tetroxide in cacodylate buffer containing 1.6% potassium ferricyanide. The discs were then washed three times with cacodylate buffer, three times with water, incubated with 0.5% uranyl acetate for 30 min (in the dark), and washed again with water. The samples were dehydrated in a graded series of ethanols while progressively lowering the temperature from 4°C to −40°C then embedded in epon resin. After peeling off the aclar, 60-nm sections were cut and viewed in an electron microscope (FEI Tecnai 12; Phillips) at 120 kV, images were captured with a digital camera at 6,800 magnification. Image is similar to Fig 6C left panel (CIL# 13758) of J Cell Biol. 2010. 191: 1173-1187.