TEM USER TRAINING

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 摘要： 该资源目前有三台可供使用的电子显微镜：JEM-4000FX(“IVEM”)、AEI EM-700万伏(“HVEM”)和FEI F-20型电子显微镜。在F-20上进行了单粒子低温电磁扫描和部分层析成像，在JEM-4000上进行了大部分低温层析数据采集，在高压电子显微镜上进行了大部分厚塑性截面层析数据采集。 在所有情况下，项目都是事先与用户讨论的，以确保标本和显微镜条件适合手头的研究。在层析、单粒子或微量分析应用的情况下，还提供了对结果的解释和进一步处理的帮助。还提出了为F-20或JEM-4000制备深度冷冻样品，以及如何为HVEM染色厚片的建议。 邢蒙负责F-20，大卫·巴纳德负责Hvem，迈克尔·马尔科负责JEM-4000。这些人员对所有内部和外部用户进行初始培训。克里斯蒂安·伦肯又在F-20上做了一些训练，大卫·曼库斯将接手，因为克里斯蒂安·伦肯已经离开了RVBC。 F-20有一个现代的用户界面，它可以设置为相对缺乏经验的显微镜专家使用，唯一的特殊预防措施是加载样本。用户将需要学会SerialEM收集断层扫描倾斜系列，并将需要学习如何识别好的样本和好的图像为单粒子冷冻-EM JEM-4000拥有广泛的系统，包括低剂量半自动断层扫描、EFTEM和EELS、STEM和X射线显微分析。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，则需要根据用户的经验进行广泛的培训。对于涉及高级技术的小项目，对于没有经验的用户，显微镜由M.Marko操作，用户在一旁观看。除了有经验的内部用户外，M.Marko为每个用户适当地设置显微镜，并更换标本。 在HVEM培训期间，David Barnard与用户坐在一起，指导如何倾斜标本、使用电视系统进行对焦、拍照等。然后，他负责在出现任何问题时帮助用户，并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.