TEM USER TRAINING

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， 因此可以在其他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： 该资源目前有三台电子显微镜可供用途：JEM-4000 FX（“IVEM”）、AEI EM-7百万伏TEM（“HVEM”）和FEI F-20 TEM。 单粒子冷冻EM工作和一些层析成像是在F-20上完成的，大部分冷冻层析成像数据收集是在JEM-4000上完成的，大部分厚塑料切片层析成像数据收集是在HVEM上完成的。 在所有情况下，项目事先与用户讨论，以确保标本和显微镜条件将适合手头的研究。 在层析成像、单粒子或微量分析应用的情况下，还可以对结果进行解释和进一步处理。 还提供了建议，在准备快速冷冻标本的rhe F-20或JEM-4000，并在如何染色厚切片的HVEM。 邢萌负责F-20，大卫巴纳德负责HVEM，迈克尔马尔科负责JEM-4000。 这些人为所有内部和外部用户进行初始培训。 克里斯蒂安·伦肯已经在F-20上进行了一些训练，大卫·曼库斯将接任，因为克里斯蒂安·伦肯已经离开了RVBC。 F-20有一个现代化的用户界面，它可以设置为一个相对缺乏经验的显微镜使用，唯一的特殊预防措施是加载标本。 用户将需要被教导SerialEM收集断层倾斜系列，并需要学习如何识别单粒子冷冻EM的良好标本和良好图像 JEM-4000具有广泛的系统，包括低剂量半自动断层扫描、EFTEM和EELS、STEM和X射线微量分析。 对于显微镜的普通使用，只需要很短的培训时间，但对于特殊模式和操作，需要根据用户的经验进行广泛的培训。 对于涉及先进技术的短期项目，为没有经验的用户，显微镜是由M。除了有经验的内部用户，M。Marko为每个用户适当设置显微镜，并更换样品。 在HVEM培训课程期间，大卫巴纳德坐在用户旁边，指导用户如何倾斜标本、使用电视系统进行对焦、拍照等。如果出现任何问题，他负责协助用户更换标本和冲洗底片。

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 Resource currently has three electron microscopes for use: The JEM-4000FX ("IVEM"), the AEI EM-7 million-volt TEM ("HVEM"), and the FEI F-20 TEM. Single-particle cryo-EM work, and some tomography, is done on the F-20, most cryo-tomographic data collection is done on the JEM-4000, and most thick-plastic-section tomographic data collection is done on the HVEM. In all cases, the project is discussed beforehand with the user to insure that the specimen and microscope conditions will be appropriate for the study at hand. Assistance is also given with interpretation of results and further processing in the case of tomography, single-particle, or microanalysis applications. Advice is also given in preparing plunge-frozen specimens for rhe F-20 or JEM-4000, and in how to stain thick sections for the HVEM . Xing Meng is responsible for the F-20, David Barnard for the HVEM and Michael Marko for the JEM-4000. These individuals do the initial training for all users, both internal and external. Christian Renken has in turn done some training on the F-20, and David Mankus will take over, now that Christian Renken has left the RVBC. The F-20 has a modern user interface, and it can be set up for a relatively inexperienced microscopist to use, the only special precaution being loading of the specimen. The user will need to be taught SerialEM to collect tomographic tilt series, and will need to learn how recognize good specimens and good images for single-particle cryo-EM The JEM-4000 has a wide range of systems, including low dose semi automated tomography, EFTEM and EELS, STEM, and X ray microanalysis. For ordinary use of the microscope, only a short training period is required, but for special modes and operations extensive training is required, depending on the experience of the user. For short projects involving advanced techniques for inexperienced users, the microscope is operated by M. Marko with the user looking on. Except for experienced internal users, M. Marko sets up the microscope appropriately for each user, and changes the specimens. During HVEM training sessions, David Barnard sits with the user, and gives instructions on how to tilt the specimen, use the TV system for focusing, take pictures, etc. He is then responsible for assisting the user should any problems arise, and for changing specimens and developing negatives.