AUTOMATED MICROSCOPY FOR ELECTRON TOMOGRAPHY
AUTOMATED MICROSCOPY FOR ELECTRON TOMOGRAPHY
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DOI：
10.1016/0304-3991(92)90016-d
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发表时间：
1992-10-01
期刊：
影响因子：
2.2
通讯作者：
AGARD, DA
中科院分区：
文献类型：
--
作者：
KOSTER, AJ;CHEN, H;AGARD, DA
描述了用于单颗粒三维重建的层析倾斜系列数据的自动收集的仪器和方法。该系统由Philips EM 430 TEM、Gatan 673冷却慢扫描CCD相机和Philips C400显微镜计算机控制单元组成。数据采集过程包括CCD摄像机上图像的直接数字记录和（a）由倾斜样品引起的图像偏移、（B）散焦变化和（c）样品的同心高度位置的自动测量和校正。实验说明自动数据收集的可能性和局限性。在30 k放大倍率下的数据收集表明，与TEM的手动操作相比，样品对光束的暴露时间减少了10-100倍。
Instrumentation and methodology for the automatic collection of tomographic tilt series data for the three-dimensional reconstruction of single particles is described. The system consists of a Philips EM 430 TEM, with a Gatan 673 cooled slow-scan CCD camera and a Philips C400 microscope computer control unit attached. The procedure for data collection includes direct digital recording of the images on the CCD camera and the automatic measurement and correction of (a) image shifts resulting from tilting the specimen, (b) variation of defocus and (c) the eucentric height position of the specimen. Experiments are described illustrating the possibilities and limitations of automatic data collection. Data collection at a magnification of 30k shows that the exposure time of the specimen to the beam is reduced by a factor of 10-100 compared to manual operation of the TEM.