Light microscopy The facility is well supported by the Office of the Scientific Director. The MIC is equipped with six modern confocal microscopes, each optimized for certain applications: 1) A Zeiss LSM 710 inverted for high-resolution confocal imaging of fixed specimen and live cells. 2) A Zeiss LSM 780 for challenging specimens that require both high resolution and high sensitivity. 3) A Nikon Spinning Disk / Total Internal Reflection Fluorescence (TIRF) hybrid microscope for high-speed confocal imaging or selective recording of membrane-bound events in live cells (TIRF). 4) A Zeiss LSM 880 2-photon confocal for thick tissues and live animals. 5) A Zeiss 800 optimized for advanced tiling experiments and, 6) A Zeiss 880 AiryScan with twice the spatial resolution of conventional microscopes without the need for special dyes or protocols. Several conventional (wide-field) light microscopes provides imaging modalities such as transmission (visible stains), large-scale tiling of tissue slices, high-speed phase contrast and DIC, and large specimens. High-end computer workstations with imaging software (Zeiss Zen, Nikon Element, Bitplane Imaris, SVI Hyugens and ImageJ) are also available. After an initial orientation where their project is researched by the staff and the best approach is decided upon, users receives hands-on training on the equipment and / or software best suited to their goals, followed by continuous support when required. Once image acquisition is complete, the staff devise solutions and train users on how to extract usable data from their images. Additional training and support is offered to the community in different ways: 1) On-site assistance and training on equipment owned by individual investigators. 2) An extensive yearly workshop covering light and electron microscopy, image analysis and sample processing. 3) The MIC staff is volunteering time to teach FAES classes. 4) The facility organizes frequent on-campus demonstrations of new instruments and software by vendors in a dedicated space. These equipment demonstrations are open to the entire NIH community. The MIC has a total of 291 registered users in 65 laboratories, and provides almost 10,000 hours of instrument time during the last fiscal year. NICHD used 80% of our resources, NINDS used 15%, and the remaining 5% went to other Institutes (NIBIB, NIA and NIMH). 20% of this instrument time was used by the MIC staff for training, internal projects and pilot experiments. Electron microscopy The electron microscopy branch of the facility processes specimens from start to finish: fixation, embedding, semi-thin and ultra-thin sectioning, staining and imaging on the JEOL 1400 transmission electron microscope. Because of the labor involved, the volume is necessarily smaller than the light microscopy branch where end users do their own processing. In the past 12 months, Mr. Dye processed a total of 130 samples, including 2 labor-intensive immuno-EM studies. The JEOL 1400 electron microscope continues to be available on the calendar for trained users. After the MIC, the unit of Dr. Zimmerberg is the major user of that instrument. Tissue preparation Mrs. Holtzclaw provides sample processing training and services for light and electron microscopy applications, including rodent perfusion, cryo-preservation, cryo-sectioning, immuno-fluorescence and tissue clearing. She dedicated a significant amount of time optimizing fixation and preservation techniques for electron microscopy studies of the central and peripheral nervous system. During the past 12 months, she provided service to Drs. Dever, Dufau, Fields, Hoffman, Klein, Le Pichon, Loh, Marini, McBain, Pfeifer, Sackett, Stojilkovic and Stopfer (NICHD), together with Drs. Mankodi, Roche, Youle and Ward (NINDS); Penzo, Plenz (NIMH); and Chen (NIBIB). Mrs. Holtzclaw also engaged in collaborative efforts with Drs. David Klein (published in PLOS One in October 2018) and Katherine Roche (Journal of Biological Chemistry, June 2019). An ongoing collaboration with Dr. Richard Youle to study the accumulation of ubiquitinated protein aggregates in brain and liver of a TAX1BP1 knock-out mouse is underway. Publications Since its inception in 2004, the work carried out in the MIC has been included in more than 185 publications. For a complete list, head to: https://science.nichd.nih.gov/confluence/display/mic/Publications
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