NICHD Microscopy and Imaging Core
- Vincent Schram, PhD, Acting Executive Director
- Tamás Balla, MD, PhD, Scientific & Administrative Director
- Ling Yi, PhD, Staff Scientist
- Louis (Chip) Dye, BS, Research Assistant
The mission of the NICHD Microscopy and Imaging Core (MIC) is to provide service in four different areas:
- (1) histology and sample preparation for light and electron microscopy;
- wide-field and confocal light microscopy;
- transmission electron microscopy (TEM); and
- image analysis and data extraction.
The Facility operates as a ‘one-stop shop,’ where investigators can, with a minimum of effort, go from their scientific question to the final data.
Mode of operation
Located on the ground floor of the Porter Building (Building 35A), the MIC is accessible 24/7, and users can reserve time on each microscope by using an online calendar The facility is available free of charge to all NICHD investigators and, resources allowing, to anyone within the Porter building. The facility is supported by the Office of the Scientific Director, NICHD.
Vincent Schram is the point person for light microscopy and data analysis and is also the team lead. Ling Yi is in charge of the histology/sample preparation unit. The Electron Microscopy (EM) branch of the Facility is staffed by Chip Dye. Tamás Balla is the scientific and administrative director of the core.
The MIC has an open-door policy with the NINDS Light Imaging Facility (LIF) in building 35. The two cores freely exchange users, share equipment, and trade support. Although not officially sanctioned, this mode of operation provides extended support hours, wider expertise, and access to more equipment than each Institute could afford on its own.
The MIC serves over 300 registered users in 68 laboratories. NICHD uses 80% of the facility's resources, NINDS 15%, and other Institutes (NIBIB, NIA, and NIMH) the remaining 5%.
Light microscopy
The MIC is equipped with six confocal microscopes, each optimized for certain applications:
- a Zeiss LSM 710 inverted for high-resolution confocal imaging;
- an LSM 900 equipped with an AiryScan detector;
- a Zeiss 800 optimized for advanced tiling experiments;
- a Zeiss 880 AiryScan with higher spatial resolution;
- the Zeiss LSM 880 2-photon confocal was donated to an NICHD investigator, and replaced with a Leica Stellaris FLIM/STED system; and
- a Nikon Spinning Disk/Total Internal Reflection Fluorescence (TIRF), equipped with advanced rotating TIRF capabilities.
The facility acquired a second automated slide scanner, a Zeiss Axioscan 7, to complement the existing Axioscan Z1. Both scanners are heavily used and free up hundreds of personnel hours for several research groups in the DIR. We continue to operate a high-end wide-angle fluorescence microscope for non-confocal imaging.
We provide image analysis services based on ImageJ, Zeiss Zen, Nikon Elements, and Bitplane Imaris. The MIC is not connected to the NIH network, so that users must move their data on removable drives.
The light microscopy branch of the MIC continues to rely on the following mode of operation: after an initial orientation, during which their project is researched by the staff and the best approach is decided upon, users receive 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 section 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 for the light microscopy branch, in which end users perform their own processing and imaging. In the past 12 months, Chip Dye processed a total of 106 samples for morphology studies.
Dye continues to operate the automatic sample preparation device from Microscopy Solutions, which allows a larger volume of specimens to be processed. John Heuser, an expert in electron microscopy, continues to use the JEOL 1400 microscope and brings his extensive experience to the MIC.
Tissue preparation
Staffed by Ling Yi, the histology/sample preparation lab is a critical component of the MIC. Nineteen users were trained in person in rodent perfusion, cryopreservation, cryo-sectioning, immunofluorescence, and RNAscope. Perfusion and cryo-sectioning services were provided to twelve laboratories. Ling Yi invested heavily in implementing RNAscope applications in the MIC, to the point where it has become routine for many of the facility's users. She is currently working on optimizing tissue clearing methods by shortening the long incubation times these techniques require. She also developed several tutorial and white papers on sample preparation, in particular a series of videos on challenging rodent dissections.
Image analysis
As mentioned above, the MIC continues to provide high-end image processing based on ImageJ, Zeiss Zen, Nikon Element, and Bitplane Imaris. For difficult cases, in which conventional processing is not suitable, the Nikon NIS-AI suite provides sophisticated tools for image restoration, segmentation, and feature extraction.
Collaborators
- John Heuser, PhD, Section on Integrative Biophysics, NICHD, Bethesda, MD
- Carolyn L. Smith, PhD, Light Imaging Facility, NINDS, Bethesda, MD
Contact
For more information, email schramv@mail.nih.gov or visit https://www.nichd.nih.gov/about/org/dir/other-facilities/cores/microscopyandimaging.
