NICHD Microscopy and Imaging Core Facility

Eunice Kennedy Shriver National Institute Of Child Health & Human Development

Investigators

Linked publications & trials

Abstract

Light microscopy The MIC is equipped with five point scanning confocal microscopes: 1) A Zeiss LSM 710 inverted for regular confocal imaging. 2) A Zeiss 900 equipped with an entry-level Airyscan detector, 3) A Zeiss 800 for tiled imaging and 4) A Zeiss 880 AiryScan with a spectral detector and a full-feature Airyscan detector. 5) The Zeiss 880 2-photon was donated to an NICHD investigator and replaced with a powerful Leica Stellaris equipped with a white light laser and capable of 3-color STED and full lifetime imaging. We also have a Nikon Spinning Disk / Total Internal Reflection Fluorescence (TIRF) instrument, equipped with advanced rotating TIRF capabilities. The facility operates several wide-field fluorescence microscopes, in particular two automatic slide scanners that are heavily used (the newer Zeiss Axioscan 7 was delivered in April 2023). For image processing and data analysis, ImageJ, Zeiss Zen, Nikon Elements, and Bitplane Imaris are available on a high-end workstation. The MIC is still cut off from the NIH network, a sad situation that seems to be permanent and is adversely affecting our operations. The light microscopy branch of the MIC continues to operate in the same semi-assisted way: after an initial orientation where their project is researched by the staff and the best approach is agreed 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. Due to their complexity, the two slides scanners are operated solely by Dr. Schram, a free service that proved very popular and is heavily used. Tissue preparation Dr. Ling continues to provide histology and sample preparation services for the Core. During the past 12 months, 93 users were trained in-person in rodent perfusion, tissue cryo-preservation, tissue sectioning (cryo, microtome and vibratome sectioning), immunofluorescence and RNAscope staining. Furthermore, perfusion, cryo-sectioning and immunofluorescence services were rendered to 20 research groups both from NICHD and from other Institutes that included NIAAA, NIMH, NCI and NINDS. The MIC has gained procedural expertise in RNAscope staining to the point where we can assist users on a routine basis. Dr. Ling also optimized tissue clearing methods by shortening the long incubation time they typically require. The technique has been adopted for immunofluorescence by researchers within NICHD and NINDS. Dr. Ling is in the process of setting up the paraffin-embedded tissue processing equipment the MIC received recently, although there has been some delays in obtaining the required space. 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 125 samples for morphology studies and one sample for immuno EM. The EM lab continues to rely on the Microscopy Innovations' mPrep ASP-2000 specimen processor, a device that reduce sample processing times up to 80%. 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. Publications Since its inception in 2004, the work carried out in the MIC has been included in more than 278 publications. For a complete list, head to: https://www.nichd.nih.gov/about/org/dir/other-facilities/cores/microscopyandimaging/publications