NICHD Microscopy and Imaging Core Facility

Eunice Kennedy Shriver National Institute Of Child Health & Human Development

Investigators

Linked publications, trials & patents

Abstract

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. In addition, the Core just received two new confocal systems: 5) A Zeiss 800 optimized for advanced tiling experiments and, 6) A Zeiss 880 Airy, which double the spatial resolution of conventional microscopes without using 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. At 11,933 instrument hours, overall usage has grown since last fiscal year, although this figure now includes the animal perfusion station and the JEOL electron microscope. 60% of that usage is coming from NICHD investigators, most of them within the Porter building. The MIC used an additional 20% of these resources for training, internal projects and pilot experiments. Other Institutes, predominantly NINDS, used the remaining 20%. Usage of each confocal system was uneven, with the 710 and 780 providing being most heavily used. However, it is worth noting the Zeiss 800 has only been in service since January 2018, and the 880 Airy since April 2018. Electron microscopy The electron microscopy branch of the facility processes specimens from start to finish: fixation, embedding, cutting, ultra-fine 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 67 samples: 53 from NICHD investigators, 2 for MIC internal test projects, and 12 from other Institutes. While this number represents a reduction compared to last years figure, it should be stressed it includes technically challenging and labor-intensive projects for Drs. Balla and Serpe. The JEOL 1400 electron microscope continues to be available on the calendar for trained users. After the MIC, Dr. Zimmerbergs unit is the major user of that instrument. Tissue preparation Mrs. Holtzclaw continues to provide sample processing training and services to the facilitys users, both for light and electron microscopy applications. She dedicates a significant amount of time to training users in different techniques such as rodent perfusion, cryopreservation, cryosectioning, immunofluorescence and tissue clearing (21 users trained in the past 12 months). She processed samples and acquired images for Drs. Balla, Buonanno, Fields, Heuser, Hoffman, Klein, Le Pichon, Loh, Marini, Ozato, Petros, Porter, Sackett, Stojilkovic and Stopfer. She has also provided training and services to Drs. Gordon, Mankodi, Youle (NINDS), Penzo, Plenz, Usdin (NIMH), Chen (NIBIB) and Danner (CC). A collaborative endeavor with Dr. David Klein was completed in July 2018, characterizing pineal cell types for which genes of interest had been documented by RNA-seq. The manuscript is currently being prepared for publication. Ms. Holtzclaw has also joined a collaborative effort with the NINDS laboratory of Dr. Richard Youle to study the accumulation of ubiquitinated protein aggregates in brain and liver of a TAX1BP1 knock-out mouse. 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