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Marker-free method for accurate alignment between correlated light cryo-light and electron cryo-microscopy data using sample support features

机译:使用样品支持功能的无标记方法可在相关光冷冻光和电子冷冻显微镜数据之间精确对准

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摘要

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.
机译:原则上,将荧光显微镜与电子断层扫描相结合,可以在细胞环境中实现标记的大分子组装体和结构特征的空间定位。为了在两个不同的成像模态之间进行精确的定位和比例积分,需要精确的对准程序。在这里,我们描述了一种无标记方法,该方法可利用光或低温荧光显微镜和电子低温显微镜来对齐图像,该方法利用了样品支撑特征,即碳膜中的孔。我们发现,根据孔中心坐标的预测误差判断,该方法的精度优于100 nm。

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