相关低温光激活定位显微镜技术与低温电子断层扫描技术
Correlated cryogenic photoactivated localization microscopy and cryo-electron tomography.
作者信息
Chang Yi-Wei, Chen Songye, Tocheva Elitza I, Treuner-Lange Anke, Löbach Stephanie, Søgaard-Andersen Lotte, Jensen Grant J
机构信息
1] Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, California, USA. [2] Howard Hughes Medical Institute, Pasadena, California, USA.
Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, California, USA.
出版信息
Nat Methods. 2014 Jul;11(7):737-9. doi: 10.1038/nmeth.2961. Epub 2014 May 11.
Cryo-electron tomography (CET) produces three-dimensional images of cells in a near-native state at macromolecular resolution, but identifying structures of interest can be challenging. Here we describe a correlated cryo-PALM (photoactivated localization microscopy)-CET method for localizing objects within cryo-tomograms to beyond the diffraction limit of the light microscope. Using cryo-PALM-CET, we identified multiple and new conformations of the dynamic type VI secretion system in the crowded interior of Myxococcus xanthus.
冷冻电子断层扫描（CET）能够在接近天然状态下以大分子分辨率生成细胞的三维图像，但识别感兴趣的结构可能具有挑战性。在此，我们描述了一种相关的冷冻光激活定位显微镜（cryo-PALM）-CET方法，用于在冷冻断层图像中定位物体，分辨率超越了光学显微镜的衍射极限。使用冷冻光激活定位显微镜-冷冻电子断层扫描（cryo-PALM-CET），我们在粘球菌拥挤的内部识别出了动态VI型分泌系统的多种新构象。
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