NICHD Microscopy and Imaging Core
- Vincent Schram, PhD, Biologist
- Louis (Chip) Dye, BS, Research Assistant
- Lynne A. Holtzclaw, BS, Research Assistant
- Sara Felsen, BS, MS, Postbaccalaureate Fellow
The mission of the NICHD Microscopy and Imaging Core (MIC) is to provide service in four different areas: (1) sample preparation for light and electron microscopy studies; (2) wide-field and confocal light microscopy; (3) transmission electron microscopy (TEM); and (4) image analysis and data extraction. The Facility is operated as a 'one-stop shop,' where investigators can, with minimum 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.
Vincent Schram is the point person for light microscopy and data analysis. The EM branch of the Facility is staffed by Chip Dye, and Lynne Holtzclaw is in charge of sample preparation (histology). Chip Dye and Lynne Holtzclaw report to Vincent Schram, who serves as acting director under the management of Chris McBain (NICHD). Tamás Balla (NICHD) acts as scientific advisor for the Facility.
The MIC has an open-door policy with the NINDS Light Imaging Facility (LIF, Building 35), where the two cores freely exchange users, equipment, and 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 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:
- Zeiss LSM 710 inverted for high-resolution confocal imaging;
- Zeiss LSM 780 with a spectral detector;
- Nikon Spinning Disk/Total Internal Reflection Fluorescence (TIRF) hybrid microscope for high-speed confocal imaging or recording of membrane-bound events in live cells (TIRF);
- Zeiss LSM 880 2-photon confocal for thick tissues and live animals;
- Zeiss 800 optimized for advanced tiling experiments;
- Zeiss 880 AiryScan with higher spatial resolution.
Several conventional (wide-field) light microscopes provide imaging modalities such as transmission (visible stains), large-scale tiling of tissue slices, high-speed phase contrast and differential interference contrast (DIC), and large specimens.
After an initial orientation, during which the staff research the project and decide on the best approach, 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 for the light microscopy branch, where end users do their own processing. In the past 12 months, Chip Dye processed a total of 145 samples for morphology studies.
Tissue preparation
Lynne Holtzclaw provided sample processing training and services for light and electron microscopy. Thirteen users were trained in-person in rodent perfusion, cryopreservation, cryosectioning, immunofluorescence and tissue clearing. With the assistance of Sara Felsen, Lynne Holtzclaw also dedicated a significant amount of time to bring RNAScope methodologies to the Core, allowing users to conduct investigations in the histology suite. NICHD users include Tamás Balla, Peter Basser, Sergey Bezrukov, Andres Buonanno, Leonid Chernomordik, Robert Crouch, Douglas Fields, Dax Hoffman, David Klein, Claire Le Pichon, Chris McBain, Karl Pfeifer, Forbes Porter, Dan Sackett, Yun-Bo Shi, Stanko Stojilkovic, and Marc Stopfer. She also provided assistance to the laboratories of Miguel Holmgren, Katherine Roche, Richard Youle, and Michael Ward (NINDS); Dietmar Plenz (NIMH); and Ellen Sidransky (NHGRI).
A collaboration with the laboratory of Richard Youle to study the accumulation of ubiquitinated protein aggregates in the brain and liver of a TAX1BP1 knock-out mouse was completed. We initiated a developmental rat pineal study, in collaboration with David Klein, with plans to probe samples using the RNAScope methodology. Lynne Holtzclaw is also working with the McBain Lab to investigate, via RNAscope, the expression of a potassium channel subunit in human parvalbumin (PV) interneurons.
Image analysis
High-end computer workstations with imaging software (Zeiss Zen, Nikon Element, Bitplane Imaris, SVI Hyugens and ImageJ) are also available.
Image processing based on neural networks (Artificial Intelligence or AI) is fast becoming a remarkably powerful tool for image restoration, segmentation, and resolution improvement. The MIC has been actively looking at AI–powered solutions for image restoration and segmentation. The Nikon NIS-AI suite, an advanced software for noise removal and segmentation not possible with conventional methods, was purchased and is now available in the core.
Collaborators
- Tamás Balla, PhD, Section on Molecular Signal Transduction, NICHD, Bethesda, MD
- David C. Klein, PhD, Scientist Emeritus, NICHD, Bethesda, MD
- Carolyn L. Smith, PhD, Light Imaging Facility, NINDS, Bethesda, MD
- Richard J. Youle, PhD, Biochemistry Section, NINDS, Bethesda, MD
Contact
For more information, email schramv@mail.nih.gov or visit http://mic.nichd.nih.gov.
