去顶细胞的相关荧光超分辨率定位显微镜和铂复型电子显微镜技术
Correlative Fluorescence Super-Resolution Localization Microscopy and Platinum Replica EM on Unroofed Cells.
作者信息
Sochacki Kem A, Taraska Justin W
机构信息
National Heart Lung and Blood Institute, National Institutes of Health, Bldg. 50, RM 3312, 50 South Drive, Bethesda, MD, 20892, USA.
出版信息
Methods Mol Biol. 2017;1663:219-230. doi: 10.1007/978-1-4939-7265-4_18.
Abstract
Platinum replicas of unroofed mammalian cells can be imaged with a transmission electron microscope (TEM) to produce high contrast, high resolution images of the structure of the cytoplasmic side of a plasma membrane. A complementary approach, super-resolution fluorescence localization microscopy, can be used to localize labeled molecules with better than 20 nm precision in cells. Here, we describe a correlative method that couples these two techniques and produces images where localization microscopy data can be used to highlight specific proteins of interest within the structural context of the platinum replica TEM image. This combined method is uniquely suited to investigate the nanometer-scale structural organization of the plasma membrane and its associated organelles and proteins.
摘要
无顶哺乳动物细胞的铂复制品可以用透射电子显微镜（TEM）成像，以生成质膜细胞质侧结构的高对比度、高分辨率图像。一种互补的方法，即超分辨率荧光定位显微镜，可以用于在细胞中以优于20纳米的精度定位标记分子。在这里，我们描述了一种关联方法，该方法将这两种技术结合起来，并生成图像，在这些图像中，定位显微镜数据可用于在铂复制品TEM图像的结构背景下突出感兴趣的特定蛋白质。这种组合方法特别适合于研究质膜及其相关细胞器和蛋白质的纳米级结构组织。
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