NICHD Microscopy and Imaging Core Facility

Eunice Kennedy Shriver National Institute Of Child Health & Human Development

Investigators

Linked publications, trials & patents

Abstract

COVID pandemic Following lower infection numbers in Montgomery County, activity in the MIC has markedly increased since the last fiscal year. For the first time since 2019, we were able to deliver our 2-week comprehensive workshop on microscopy using an hybrid in-person / remote format. The number of on-site equipment demonstrations significantly increased, with a particular involvement of Nikon and Leica. The complete MIC staff is on campus full time, although the general scientific activity has not reached pre-pandemic levels yet. 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) The Nikon Spinning Disk / Total Internal Reflection Fluorescence (TIRF), equipped with advanced rotating TIRF capabilities. 3) A Zeiss LSM 880 2-photon confocal for thick tissues and live animals. 4) A Zeiss 800 optimized for advanced tiling experiments and, 5) A Zeiss 880 AiryScan with higher spatial resolution. The older Zeiss LSM 780 was retired in July 2022 and replaced with a modern LSM 900 equipped with an AiryScan detector. The fully automatic Zeiss Axioscan Z1 slide scanner was heavily used in FY 2022, freeing up hundreds of personnel hours for several research units in the DIR. We also completed a high-end wide-angle fluorescence microscope to complement our equipment base for non-confocal microscopy imaging. High-end computer workstations with imaging software (Zeiss Zen, Nikon Elements, Bitplane Imaris, SVI Hyugens and ImageJ) continue to be used for data extraction. Unfortunately, the MIC started to be handicapped by acute network connectivity issues in June 2022. This crippling problem shows no sign of being resolved. The light microscopy branch of the MIC continues to rely on a the same mode of operation. 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 and imaging. In the past 12 months, Mr. Dye processed a total of 148 samples for morphology studies. An automatic sample preparation device from Microscopy Solutions was acquired and installed, allowing to increase the volume of specimens processed. Dr. John Heuser, a seasoned expert in electron microscopy, continues to use the JEOL 1400 microscope and interface with Mr. Dye, bringing his extensive experience to the MIC. Tissue preparation Mrs. Lynne Holtzclaw, the point person for histology services in the MIC for many years, retired in August 2021 (we wish her a happy retirement!). The MIC was able to hire a talented staff scientist, Dr. Ling Yi, to take over this role. Dr. Ling has done a remarkable job insuring a smooth transition from Mrs. Holtzclaw, and is now providing sample processing and histology services for light and electron microscopy. 22 users were trained in-person in rodent perfusion, cryopreservation, cryosectioning, immunofluorescence and RNAscope. Perfusion and cryosectioning services were provided to 8 laboratories. NICHD users include Drs. Buonanno, D'Souza, Fields, Hoffman, Le Pichon, Pfeifer and Weinstein. Users from other Institutes include Drs. Joseph (NIAAA) and Penzo (NIMH). Dr. Ling heavily invested in implementing RNAscope applications in the MIC, to the point where it has become routine to many of the facility's users. She is currently working on optimizing tissue clearing methods by shortening the long incubation time these techniques require. Publications Since its inception in 2004, the work carried out in the MIC has been included in more than 250 publications. For a complete list, head to: https://www.nichd.nih.gov/about/org/dir/other-facilities/cores/microscopyandimaging/publications