Cédric S. Asensio, Daniel W. Sirkis, Robert H. Edwards (2011) CIL:13758, Rattus norvegicus, pheochromocytoma, adrenal gland cell. CIL. Dataset

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 Fig 6C left panel and is control for Fig 6C right panel (CIL# 13764) of J Cell Biol. 2010. 191: 1173-1187.

电镜图像展示经对照siRNA转染的PC12细胞。PC12细胞接种于涂有聚赖氨酸的Pella aclar薄膜圆盘上，经50 nM的对照siRNA转染两次，并使用不含钙镁的PBS溶液中的2.5%戊二醛固定。圆盘经0.1 M的钠草酸盐缓冲液（pH 7.2）清洗三次，并在冰上用含有1.6%铁氰化钾的草酸盐缓冲液中的1%四氧化锇进行后固定30分钟。随后，圆盘用草酸盐缓冲液清洗三次，再用水清洗三次，并在暗处用0.5%醋酸铀酸酯孵育30分钟，最后再次用水清洗。样品在逐渐降低温度从4°C至-40°C的过程中，用分级乙醇进行脱水，然后浸入环氧树脂中。去除aclar膜后，切割出60纳米厚的切片，并在120 kV的FEI Tecnai 12（Philips）电子显微镜下观察，使用数字相机在6,800倍放大下捕捉图像。图像为J Cell Biol. 2010年第191卷第1173-1187页的图6C左侧面板，是图6C右侧面板的对照（CIL# 13764）。