TOMOGRAPHY TRAINING OF VISITORS TO THE LAB

实验室访客的断层扫描培训

• 批准号: 8170826
• 负责人: CINDY L SCHWARTZ
• 金额: $ 2.49万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170826
• 关键词: Amber; Antibodies; Austria; Computer Retrieval of Information on Scientific Projects Database; Computer software; Computer-Assisted Image Analysis; Cytoskeleton; Data; Data Set; Epithelial Cells; Fishes; Fission Yeast; Freezing; Funding; Germany; Grant; Hair Cells; Image; Image Analysis; Institutes; Institution; Israel; Laboratories; Learning; Mediating; Meiosis; Methods; Microscope; Microtubules; Modeling; Neurons; Plants; Plastic Embedding; Plastics; Preparation; Programmed Learning; Research; Research Personnel; Resolution; Resources; Rotation; Series; Source; Specimen; Stereocilium; Students; Training; United States National Institutes of Health; Universities; Visit; Wolves; Yeasts; coping; graduate student; neonatal Fc receptor; programs; reconstruction; spindle pole body; tomography

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. This year our facility hosted a total of 15 groups of visitors, each of whom spent one week or more to learn various methods of specimen preparation, and/or to collect tilt series data and learn computer-assisted image analysis. This year, 7 visitors and students came specifically to collect cryo-tilt series and to learn the cryotomography methods developed in our lab. Herbert Poltzhofer and Guenter Resch (IMBA Vienna, Austria) visited the lab on two separate occasions to collect tilt series of frozen-hydrated whole mounts of fish keratocytes. Daniela Nicastro (Brandeis Univ.) spent three days collecting tilt series of isolated, plunge frozen microtubules decorated with klp 6. Dilem Hizlan (EMBL, Heidelberg, Germany) spent a week in the lab imaging isolated, plunge frozen microtubules with centralspindlin. In addition, three graduate students in MCDB, (Amber Rex, Julia Cope, Ian Ross, and Nesia Zurek) did rotations in the 3D lab to learn cryomicroscopy, cryotomography and image analysis. A fourth rotation student, Ian Ross, learned tomography of plastic-embedded meiotic yeast spindle pole bodies. Eight visitors came to collect data from plastic sections on the F30 microscope and/or to learn the various methods of 3D reconstruction using the IMOD software package. Each group was able to master aspects of the IMOD software package and has gone on to install the software in their own laboratories. Dr. Wanzhong He (CalTech University) spent 5 days collecting tilt series data on the F30 and learning the programs to study FcRn-mediated antibody transport across epithelial cells. Details of this project can be found in the Research Highlights. Dr. Hildur Palsdottir (Lawrence Berkeley Lab) spent three days collecting tilt series data of hair cell stereocilia. Helio Roque (EMBL, Heidelberg), collected montage tilt series data on the F20 and learned the IMOD modeling program and programs for tomographic reconstruction of the microtubule cytoskeleton in S. pombe. Mitusuko Hayashi (Yale University) collected tilt series data on the F30 microscope of neurons. Endre Majorovits (Oxford University) spent a week collecting tilt series data and learning the image analysis software for tomographic reconstruction and modeling. Ichirou Karahara (Univ. Toyama) collected tilt series data on the F30 microscope of plant cytoskeleton. Sharon Wolf (Weizmann Institute, Israel) spent a day with Cindi Schwartz learning details of the Serial EM image acquisition software for high-resolution data sets.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 今年，我们的设施总共接待了 15 组参观者，每组参观者花费一周或更长时间来学习各种标本制备方法，和/或收集倾斜系列数据并学习计算机辅助图像分析。 今年，7位参观者和学生专门来收集冷冻倾斜系列并学习我们实验室开发的冷冻断层摄影方法。 Herbert Poltzhofer 和 Guenter Resch（IMBA 维也纳，奥地利）两次访问该实验室，收集倾斜系列的冷冻水合整批鱼类角膜细胞。 Daniela Nicastro（布兰代斯大学）花了三天时间收集倾斜系列的用 klp 6 装饰的分离的、插入冷冻的微管。Dilem Hizlan（EMBL，德国海德堡）花了一周的时间在实验室对带有中枢轴蛋白的分离的、插入冷冻的微管进行成像。此外，MCDB 的三名研究生（Amber Rex、Julia Cope、Ian Ross 和 Nesia Zurek）在 3D 实验室进行轮转，学习冷冻显微镜、冷冻断层扫描和图像分析。第四轮轮换的学生伊恩·罗斯（Ian Ross）学习了塑料嵌入的减数分裂酵母纺锤体的断层扫描。八位参观者前来收集 F30 显微镜上的塑料切片数据和/或学习使用 IMOD 软件包进行 3D 重建的各种方法。每个小组都能够掌握 IMOD 软件包的各个方面，并继续在自己的实验室中安装该软件。 Wanzhong He 博士（加州理工大学）花了 5 天时间收集 F30 上的倾斜系列数据，并学习了研究 FcRn 介导的抗体跨上皮细胞转运的程序。该项目的详细信息可以在研究亮点中找到。 Hildur Palsdottir 博士（劳伦斯伯克利实验室）花了三天时间收集毛细胞静纤毛的倾斜系列数据。 Helio Roque（EMBL，海德堡）收集了 F20 上的蒙太奇倾斜系列数据，并学习了 IMOD 建模程序和裂殖酵母微管细胞骨架的断层扫描重建程序。 Mitusuko Hayashi（耶鲁大学）在 F30 神经元显微镜上收集倾斜系列数据。 Endre Majorovits（牛津大学）花了一周的时间收集倾斜系列数据并学习用于断层扫描重建和建模的图像分析软件。 Ichirou Karahara（富山大学）在 F30 显微镜上收集了植物细胞骨架的倾斜系列数据。 Sharon Wolf（以色列魏茨曼研究所）花了一天时间与 Cindi Schwartz 一起学习用于高分辨率数据集的 Serial EM 图像采集软件的详细信息。