Downing Kenneth H, Mooney Paul E
Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA.
Rev Sci Instrum. 2008 Apr;79(4):043702. doi: 10.1063/1.2902853.
Electron microscopists are increasingly turning to intermediate voltage electron microscopes (IVEMs) operating at 300-400 kV for a wide range of studies. They are also increasingly taking advantage of slow-scan charge coupled device (CCD) cameras, which have become widely used on electron microscopes. Under some conditions, CCDs provide an improvement in data quality over photographic film, as well as the many advantages of direct digital readout. However, CCD performance is seriously degraded on IVEMs compared to the more conventional 100 kV microscopes. In order to increase the efficiency and quality of data recording on IVEMs, we have developed a CCD camera system in which the electrons are decelerated to below 100 kV before impacting the camera, resulting in greatly improved performance in both signal quality and resolution compared to other CCDs used in electron microscopy. These improvements will allow high-quality image and diffraction data to be collected directly with the CCD, enabling improvements in data collection for applications including high-resolution electron crystallography, single particle reconstruction of protein structures, tomographic studies of cell ultrastructure, and remote microscope operation. This approach will enable us to use even larger format CCD chips that are being developed with smaller pixels.
电子显微镜学家越来越多地转向使用工作电压为300 - 400 kV的中压电子显微镜(IVEM)进行广泛的研究。他们也越来越多地利用慢扫描电荷耦合器件(CCD)相机,这种相机已在电子显微镜上广泛使用。在某些条件下,CCD在数据质量方面比摄影胶片有所提高,并且具有直接数字读出的诸多优点。然而,与更传统的100 kV显微镜相比,IVEM上的CCD性能严重下降。为了提高IVEM上数据记录的效率和质量,我们开发了一种CCD相机系统,在电子撞击相机之前将其减速至100 kV以下,与电子显微镜中使用的其他CCD相比,在信号质量和分辨率方面都有了极大的提升。这些改进将使我们能够直接用CCD收集高质量的图像和衍射数据,从而改善包括高分辨率电子晶体学、蛋白质结构的单颗粒重建、细胞超微结构的断层扫描研究以及远程显微镜操作等应用中的数据收集。这种方法将使我们能够使用正在开发的具有更小像素的更大尺寸的CCD芯片。