ELECTRON TOMOGRAPHY TRAINING AND DISSEMINATION

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： 开始断层扫描研究的第一步是明确研究者希望从重建中了解什么。 我们可能会发现，一个简单的倾斜系列动画，或使用立体对图像与Sterecon更合适。 一旦我们同意断层摄影研究是适当的，标本厚度和放大倍数的选择必须仔细考虑。 然后训练用户用适当的电子显微镜记录倾斜系列。 在某些情况下，我们的工作人员做显微镜。 保护样品免受污染和过度辐照是很重要的。 如果图像记录在胶片上，我们会协助用户设置扫描相机，以进行图像数字化和预对准。 最终的对齐通常使用金标记作为基准标记，并使用WEB完成，然后使用SPIDER计算重建。重建的可视化是项目中最耗时的部分。 在许多情况下，我们在调查员的陪同下进行这项工作，如果调查员要独立进行这项工作，就必须接受使用SPIDER和各种可视化软件包的培训。 一些研究人员在他们自己的实验室中具有可视化功能，如果他们使用我们熟悉（或提供）的软件，我们也会提供帮助，通常是通过互联网。 有关我们使用的断层重建方法的完整文档可在SPIDER主页上找到，并定期更新。

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 first step in beginning a tomographic study is to clarify precisely what the investigator hopes to learn from a reconstruction. We may find that a simple tilt series animation, or the use of stereopair images with Sterecon is more appropriate. Once we agree that a tomographic study is appropriate, the choice of specimen thickness and magnification have to be carefully considered. The user is then trained in recording the tilt series with the appropriate electron microscope. In some cases, our staff does the microscopy. Protection of the specimen from contamination and excessive irradiation are important. If images are recorded on film, we assist the user in setting up the scanning camera for image digitization and pre alignment. The final alignment normally uses gold markers as fiducial marks and is done using WEB, then the reconstruction is computed using SPIDER. Visualization of the reconstruction is the most time consuming portion of the project. In many cases, we do this with the investigator looking on. If the investigator is to do this independently, there must be training in the use of SPIDER and various visualization packages. Some investigators have visualization capabilities in their own labs, and if they use software we are familiar with (or supply), we also provide assistance, usually over the Internet. Full documentation on the tomographic reconstruction methods we use is available on the SPIDER homepage, and is updated periodically.