TOMOGRAPHY TRAINING OF VISITORS TO THE LAB

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。 今年，我们的设施共接待了15组参观者，每个参观者都花了一周或更长的时间学习各种标本制备方法，和/或收集倾斜系列数据和学习计算机辅助图像分析。今年，7名参观者和学生专门来收集低温倾斜系列，学习我们实验室开发的低温层析成像方法。Herbert Poltzhofer和Guenter Resch(奥地利维也纳国际管理学院)在两个不同的场合访问了该实验室，收集了倾斜系列的冷冻水合完整的鱼类角质细胞。丹妮拉·尼卡斯特罗(Brandeis Univ.)Dilem Hizlan(德国海德堡EMBL)花了一周的时间在实验室对分离的、插入中央纺锤体的冷冻微管进行成像。此外，MCDB的三名研究生(琥珀·雷克斯、朱莉娅·科普、伊恩·罗斯和内西亚·祖雷克)在3D实验室进行旋转，学习冷冻显微镜、冷冻断层扫描和图像分析。第四个轮班学生，伊恩·罗斯，学习了塑料嵌入的减数分裂酵母纺锤体的断层扫描。8名参观者前来收集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。每个小组都掌握了IMOD软件包的方方面面，并在自己的实验室安装了该软件。何万忠博士(加州理工大学)花了5天时间收集F30的倾斜序列数据，并学习研究FcRN介导的抗体跨上皮细胞传输的程序。这个项目的详细情况可以在研究摘要中找到。希尔杜尔·帕尔斯多蒂尔(Lawrence Berkeley Lab)博士花了三天时间收集毛细胞立体纤毛的倾斜系列数据。Helio Roque(EMBL，Heidelberg)，收集了F20上的蒙太奇倾斜序列数据，并学习了IMOD建模程序和用于重建S.pombe微管细胞骨架的断层扫描程序。耶鲁大学的Mitusuko Hayashi收集了神经元F30显微镜上的倾斜序列数据。Endre Majorovits(牛津大学)花了一周的时间收集倾斜序列数据，并学习用于断层重建和建模的图像分析软件。Ichirou Karahara(大学TOYAMA)在F30显微镜上采集了植物细胞骨架的倾斜系列数据。Sharon Wolf(以色列魏兹曼研究所)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。

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.