Automated electron microscope tomography using robust prediction of specimen movements

A new method was developed to acquire images automatically at a series of specimen tilts, as required for tomographic reconstruction. The method uses changes in specimen position at previous tilt angles to predict the position at the current tilt angle. Actual measurement of the position or focus is skipped if the statistical error of the prediction is low enough. This method allows a tilt series to be acquired rapidly when conditions are good but falls back toward the traditional approach of taking focusing and tracking images when necessary. The method has been implemented in a program, SerialEM, that provides an efficient environment for data acquisition. This program includes control of an energy filter as well as a low-dose imaging mode, in which tracking and focusing occur away from the area of interest. The program can automatically acquire a montage of overlapping frames, allowing tomography of areas larger than the field of the CCD camera. It also includes tools for navigating between specimen positions and finding regions of interest. (C) 2005 Elsevier Inc. All rights reserved.

开发了一种新方法，可按照断层重建的要求在一系列样品倾斜角度下自动获取图像。该方法利用样品在先前倾斜角度下的位置变化来预测当前倾斜角度下的位置。如果预测的统计误差足够低，则跳过对位置或焦点的实际测量。当条件良好时，这种方法可以快速获取倾斜系列图像，但在必要时会回归到拍摄聚焦和跟踪图像的传统方法。该方法已在SerialEM程序中实现，该程序为数据采集提供了高效的环境。这个程序包括对能量过滤器的控制以及低剂量成像模式，在低剂量成像模式下，跟踪和聚焦在感兴趣区域之外进行。该程序可以自动获取重叠帧的拼接图像，从而能够对大于CCD相机视野的区域进行断层扫描。它还包括用于在样品位置之间导航以及寻找感兴趣区域的工具。（C）2005爱思唯尔公司。保留所有权利。