TOMOGRAPHY TRAINING OF VISITORS TO THE LAB

该子项目是利用 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。列出的机构是 研究中心，而研究中心不一定是研究者所在的机构。 今年，我们的设施共接待了15组参观者，每组参观者花了一周或更长时间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，有7名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扫描方法。赫伯特Poltzhofer和Guenter Resch（IMBA维也纳，奥地利）在两个不同的场合访问了实验室，以收集倾斜系列的冷冻水合的整批鱼角膜细胞。Daniela Nicastro（布兰迪斯大学）花了三天时间收集倾斜系列的用KLP 6修饰的分离的、插入的冷冻微管。Dilem Hizlan（EMBL，Heidelberg，德国）花了一周的时间在实验室中用中心纺锤体对分离的冰冻微管进行成像。此外，MCDB的三名研究生（Amber雷克斯、Julia科普、Ian Ross和Nesia Zurek）在3D实验室进行了轮换，以学习低温显微镜、冷冻断层扫描和图像分析。第四个旋转的学生，伊恩·罗斯，学习了塑料嵌入减数分裂酵母纺锤体极体的断层扫描。八位参观者前来收集F30显微镜塑料切片的数据和/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安装软件。Wanzhong He博士（加州理工大学）花了5天时间收集F30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摘要中找到。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，Heidelberg）在F20上收集了蒙太奇倾斜系列数据，并学习了IMOD建模程序和S.粟球。Mitusuko Hayashi（耶鲁大学）在F30显微镜下收集了神经元的倾斜系列数据。 Endre Majorovits（牛津大学）花了一周时间收集倾斜系列数据，并学习用于断层重建和建模的图像分析软件。Ichirou Karahara（Univ.富山）在F30显微镜上收集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串行EM图像采集软件的细节。

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.