TOMOGRAPHY TRAINING OF VISITORS TO THE LAB

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. 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.