Max Planck Institute for Medical Research, Jahnstr., 29, 69120, Heidelberg, Germany.
J Microsc. 2013 Dec;252(3):258-62. doi: 10.1111/jmi.12086. Epub 2013 Sep 24.
Serial block-face electron microscopy with focused ion beam cutting suffers from cutting artefacts caused by changes in the relative position of beam and sample, which are, for example, inevitable when reconditioning the ion gun. The latter has to be done periodically, which limits the continuous stack-acquisition time to several days. Here, we describe a method for controlling the ion-beam position that is based on detecting that part of the ion beam that passes the sample (transmitted beam). We find that the transmitted-beam current decreases monotonically as the beam approaches the sample and can be used to determine the relative position of beam and sample to an accuracy of around one nanometre. By controlling the beam approach using this current as the feedback parameter, it is possible to ion-mill consecutive 5 nm slices without detectable variations in thickness even in the presence of substantial temperature fluctuations and to restart the acquisition of a stack seamlessly. In addition, the use of a silicon junction detector instead of the in-column detector is explored.
采用聚焦离子束切割的连续块状面电子显微镜会受到由于束流和样品的相对位置变化而产生的切割伪影的影响,例如在离子枪重新调试时,这种影响是不可避免的。而后者需要定期进行,这就将连续堆叠采集的时间限制在几天以内。在这里,我们描述了一种控制离子束位置的方法,该方法基于检测穿过样品的那部分离子束(透射束)。我们发现,透射束电流随着束流接近样品而单调下降,可以用于确定束流和样品的相对位置,精度约为一纳米。通过使用该电流作为反馈参数来控制束流接近,可以在存在明显温度波动的情况下,离子铣削连续的 5nm 切片,而不会导致厚度发生可察觉的变化,并可以无缝地重新开始堆叠的采集。此外,还探索了使用硅结探测器而不是柱内探测器。