Marker-free method for accurate alignment between correlated light, cryo-light, and electron cryo-microscopy data using sample support features
Marker-free method for accurate alignment between correlated light, cryo-light, and electron cryo-microscopy data using sample support features
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DOI：
10.1016/j.jsb.2017.11.001
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发表时间：
2018-01-01
影响因子：
3
通讯作者：
Volkmann, Niels
中科院分区：
文献类型：
--
作者：
Anderson, Karen L.;Page, Christopher;Volkmann, Niels
将荧光显微镜与电子冷冻断层扫描相结合，原则上可以在细胞环境中对标记的大分子组装和结构特征进行空间定位。为了在两种不同的成像模式之间实现精确的定位和尺度集成，需要精确的校准程序。在这里，我们描述了一种无标记的方法来对准来自光或冷冻光荧光显微镜和电子冷冻显微镜的图像，该方法利用了样品支持特征，即碳膜中的空穴。从孔中心坐标的预测误差来看，该方法的精度优于100 nm。
Combining fluorescence microscopy with electron cryo-tomography allows, in principle, spatial localization of tagged macromolecular assemblies and structural features within the cellular environment. To allow precise localization and scale integration between the two disparate imaging modalities, accurate alignment procedures are needed. Here, we describe a marker-free method for aligning images from light or cryo-light fluorescence microscopy and from electron cryo-microscopy that takes advantage of sample support features, namely the holes in the carbon film. We find that the accuracy of this method, as judged by prediction errors of the hole center coordinates, is better than 100 nm.