IVEM OPERATION AND IMPROVEMENTS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 摘要： IVEM（JEOL JEM-4000 FX）主要用于TRD 1，冷冻水合标本的电子断层扫描。 IVEM配备了一个LaB 6阴极，一个5轴计算机控制的测角仪，和几个双倾斜和倾斜旋转样品台，以及Gatan单倾斜和倾斜旋转低温转移台。 2002年，我们完成了Gatan GIF 2002柱后能量过滤器的安装，该过滤器配备了2048 x2048 CCD摄像机，并安装了TVIPS FastScan电视速率CCD摄像机，用于在非常低的电子剂量下跟踪束敏感标本。 通过外部计算机控制提供自动电子断层扫描。仪器分辨率为0.14 nm（金晶格）。 STEM模式点到点分辨率为1 nm。 提供完整的分析能力，包括EELS和EDX。 上一报告所述期间的活动： 老化的Emispec ES Vision STEM成像和EDX/EELS光谱系统更换为4pi“Revolution”系统。 这是必要的，因为Emispec已不复存在，对这些装置的服务也不再可用。 我们借此机会将所有EM控制软件整合到一台新的单台计算机上。 我们安装了最新版本的Gatan和Tietz成像软件。 我们重新编写了用于收集断层倾斜系列图像的自动化系统，因为旧版本是基于Emispec系统构建的。 新版本完全独立。 它有一个与Gatan能量过滤器和CCD成像更方便的使用界面，通过内部直接命令Gatan数字显微镜。 所有的EM控制和成像现在都可以在同一个应用程序窗口中完成，数据收集所需的鼠标数量也减少了。 新软件是由迪安利斯编写的。 这样做是必要的，而不是简单地实施串行EM或其他更现代的数据收集系统，因为JEM-4000的样品台不像更现代的TEM那样同心，需要更复杂的程序来跟踪倾斜系列期间的样品。 STEM光谱成像的电子能量损失和能量色散X射线显微分析的集成，现在使用的革命系统提供的4pi公司。 4pi的总裁Scott Davilla是一名前资源工作人员，曾在我们的HVEM工作。 我们是EELS与STEM集成的beta测试站点，我们将在下一个融资周期（见下文）中将该系统与我们的新JEM-3200 FSC一起使用。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. ABSTRACT: The IVEM (JEOL JEM-4000FX) is primarily used for TRD1, electron tomography of frozen- hydrated specimens. The IVEM is equipped with a LaB6 cathode, a 5-axis computer-controlled goniometer, and several double-tilt and tilt-rotation specimen stages, as well as Gatan single-tilt and tilt-rotation cryo-transfer stages. In 2002, we completed installation of a Gatan GIF2002 post-column energy filter equipped with a 2048x2048 CCD camera, and a TVIPS FastScan TV-rate CCD camera was installed for tracking beam-sensitive specimens at very low electron dose. Automated electron tomography is provided via external computer control. The instrumental resolution is 0.14 nm (gold lattice). The STEM-mode point-to-point resolution is 1 nm. Full analytical capability is provided, including EELS and EDX. Activity in past reporting period: The aging Emispec ES Vision STEM imaging and EDX/EELS spectroscopy system was replaced with a 4pi "Revolution" system. This was necessary because Emispec no longer exists, and service on the units is no longer available. We took this opportunity to consolidate all EM control software on a new, single computer. We installed the latest versions of the Gatan and Tietz imaging software. We re-wrote our automation system for collecting tomographic tilt-series images, since the old version was built on the Emispec system. The new version is completely stand-alone. It has a much easier-to-use interface with the Gatan energy filter and CCD imaging, via internal direct commands to Gatan Digital Micrograph. All EM control and imaging can now be done from the same application window, and the number of mouse needed for data collection has been reduced. The new software was written by Dean Leith. It was necessary to do this, rather than simply implementing Serial EM or another more-modern data-collection system, because the specimen stage of the JEM-4000 is not as eucentric as that of more modern TEMs, requiring a more-complicated procedure to track the specimen during the tilt series. The integration of STEM spectrum imaging for both electron energy-loss and energy-dispersive x-ray microanalysis, is now done using the Revolution system supplied by 4pi company. The President of 4pi, Scott Davilla, is a former Resource staff member, having worked on our HVEM. We are a beta-test site for integration of EELS with STEM, and we will use this system with our new JEM-3200FSC during the next funding cycle (see below).