NICHD Microscopy and Imaging Core Facility

Eunice Kennedy Shriver National Institute Of Child Health & Human Development

Investigators

Linked publications & trials

Abstract

COVID pandemic Like every research unit on campus, the MIC continues to be affected by restrictions due to Covid19. From October 2020 until June 2021, the facility operated at half-capacity with a hard limit of two staffs and four users present at any given time. Occupancy restrictions relaxed in June 2020, allowing the facility to operate at near-normal level of staffing. The yearly MIC-organized workshop on microscopy could not be held in person and was again cancelled for 2021. The staff continue to be involved in the (online) BIOC35 microscopy course. Due to social distancing rules, there was only a handful of on-site equipment demonstrations during the past 12 months. As of this writing, the staff is back on campus full time, but most laboratories are still impacted by occupancy limits. As a result, the scientific output of the MIC has not yet rebounded to pre-pandemic levels. Light microscopy The MIC is equipped with six confocal microscopes, each optimized for certain applications: 1) A Zeiss LSM 710 inverted for high-resolution confocal imaging. 2) A Zeiss LSM 780 with a spectral detector 3) The Nikon Spinning Disk / Total Internal Reflection Fluorescence (TIRF) hybrid microscope was replaced with a new unit better suited for large-scale tiled imaging, and equipped with rotating TIRF capabilities. 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 higher spatial resolution. The MIC also received the much-anticipated Zeiss Axioscan Z1 slide scanner, which is proving tremendously useful for heavy imaging projects. High-end computer workstations with imaging software (Zeiss Zen, Nikon Elements, Bitplane Imaris, SVI Hyugens and ImageJ) are also available. Image processing based on neural networks (Artificial Intelligence or AI) is a powerful tool for image restoration, segmentation and resolution improvement. The MIC purchased a second license of the Nikon NIS-AI suite for image restoration, segmentation and multiplexing. 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. 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 110 samples for morphology studies. Tissue preparation Mrs. Holtzclaw provided sample processing and histology services for light and electron microscopy. Six users were trained in-person in rodent perfusion, cryopreservation, cryosectioning, and immunofluorescence. Perfusion and cryosectioning services were provided to 5 laboratories. NICHD users include Drs. Balla, Buonanno, Crouch, Fields, Hoffman, Le Pichon, Loh, McBain, Ozato, Pfeifer, Sackett, Stojilkovic and Stopfer. Users from other Institutes include Drs. Penzo (NIMH), Plenz (NIMH), and Cookson (NIA). A collaborative effort with Dr. Robert Crouch to investigate developmental cerebellar protein expression in the ataxia Waddle mouse was pursued. RNAScope experiments for a collaborative project with Dr. Klein were concluded with the assistance of Mrs. Felsen. These experiments targeted important developmental genes in the rat pineal gland and corroborated RNASeq data collected from dissociated pineal glands. Publications Since its inception in 2004, the work carried out in the MIC has been included in more than 200 publications. For a complete list, head to: https://www.nichd.nih.gov/about/org/dir/other-facilities/cores/microscopyandimaging/publications