Electron Microscopy Research Core

摘要 我们的目标是创建共享的电子显微镜（EM）设施，这将使UCC的神经科学界能够扩展他们的实验方法。所有拟议的勘探项目都采用了EM技术，尽管程度不同。因此，Bykhovskaia博士的项目在分析突触囊泡的组织方面严重依赖EM方法，EM设施将为此提供支持 通过帮助Bykhovskaia实验室改进EM协议，Jorquera博士的项目包含一个实验系列，在量化突触囊泡的功能池，其中EM方法被采用。 博士Jorquera和他的实验室成员将接受EM设施人员的培训，将获得设备，并将在EM设施主任的咨询和指导下进行EM实验。最后，马丁斯博士将探讨EM技术在分析神经保护药物对神经元超微结构和细胞凋亡的影响方面的能力。马丁斯博士将由EM设施主任在 实验设计和EM设施将为Martins博士提供样品制备和EM分析服务。此外，UCC神经科学社区的其他成员，包括Eaton，Skatchkov和Ferchmin博士正在规划和开发EM方法。因此，UCC的神经科学界对电子显微镜数据有着强烈的需求。这包括3D重建， 突触末端，高分辨率蛋白定位和遗传方法。重要的是，UCC的EM能力吸引了几个外部合作，包括C Tapia博士（哥伦比亚大学），N。Toni（洛桑大学），C.F.博士史蒂文斯（索尔克研究所）和罗博士（加州大学欧文分校）。重要的是，Schikorski博士的实验室最近开发了一种用于神经元电子显微镜的报告基因， 该技术目前是UCC EM设施独有的。因此，我们有能力满足这种日益增长的需求EM作为一种研究工具，并为我们的学生提供多元化背景的机会，由高素质的人才在一个强大的EM方法进行培训。为了满足这一需求，我们提出了电子显微镜研究核心（EMRC）作为当前SNRP应用的一部分。我们的目标是 EMRC将改善基础设施，简化样品制备，增加新的EM技术，并为新用户提供扩展的服务和培训。EMRC的目标是为来自不同背景的教师和学生提供高质量的培训和研究机会，并继续促进与当地和外部研究人员的有效合作。最后，EMRC的长期目标是将联合收割机电子 显微镜与超分辨率荧光显微镜，从而使我们的社区尖端的能力，神经元结构分析在纳米尺度。

Abstract Our goal is to create shared Electron Microscopy (EM) facility, which would enable the neuroscience community in the UCC to extent their experimental approaches. All the proposed exploratory projects employ the EM technique, although to a different extent. Thus, Dr. Bykhovskaia's project heavily relies on the EM approach in analyzing the organization of synaptic vesicles, and the EM facility will provide support for this project by helping the Bykhovskaia lab to improve the EM protocols. Dr. Jorquera's project contain an experimental series in quantifying functional pools of synaptic vesicles, where the EM approach is employed. Dr. Jorquera and his lab members will be trained by the EM facility personnel, will get access to the equipment, and will be consulted and guided by the EM facility Director in performing the EM experiments. Finally, Dr. Martins will explore the capabilities of the EM technique in analyzing the effect of a neuroprotective drug on neuronal ultrastructure and apoptosis. Dr. Martins will be guided by the Director of the EM facility in the experimental design, and the EM facility will provide Dr. Martins with service in the sample preparation and EM analysis. In addition, other members of the UCC neuroscience community, including Drs. Eaton, Skatchkov, and Ferchmin are in the process of planning and developing EM approaches. Thus, the neuroscience community at the UCC has a strong need for electron microscopic data. This includes 3D reconstructions of synaptic terminals, high resolution protein localization, and genetic approaches. Importantly, EM capabilities in the UCC attracted several outside collaborations, including Dr. C Tapia (Columbia University), Dr. N. Toni (Lausanne University), Dr. C.F. Stevens (The Salk Institute, and Dr. Luo (UC Irvine). Importantly, a reporter gene for electron microscopy of neurons has been recently developed in the lab of Dr. Schikorski, and this technique is currently unique to the UCC EM facility. Thus, we are well positioned to satisfy this growing need for EM as a research tool and to provide our students with diversity backgrounds with the opportunity of being trained in a powerful EM approach by highly qualified personnel. To address this need, we propose the Electron Microscopy Research Core (EMRC) as a part of the current SNRP application. Our objective for the EMRC will be to improve infrastructure, streamline sample preparation, add new EM techniques, and offer expanded services and training to new users. The EMRC Objectives are to provide high quality training and rsearch opportunities to faculty and students from diversity backgrounds and continue fostering effective collaborations with local and external researchers. Finally, the EMRC loner-term goal is to combine electron microscopy with super-resolution fiuorescence microscopy, and thus to bring to our community cutting edge capabilities for neuronal structural analysis at the nano-scale.