Infectious Diseases Collaborative Technological Research Resources- RML

National Institute Of Allergy And Infectious Diseases

Investigators

Linked publications & trials

Abstract

In FY2025, this project provides state-of-the-art research technologies that are developed, validated, and then applied in support of NIAID research. Technologies developed outside the NIH are likewise tested, evaluated, validated and, if appropriate, incorporated into the technology portfolio of the intramural program. Electron Microscopy The RML Microscopy (EM) Unit provides expertise in advanced electron microscopy (EM)-related techniques and technologies to support the ultra-structural imaging needs of the DIR scientists both in Bethesda and at the Rocky Mountain Laboratories (RML). The facility provides consultations, sample preparation, imaging, and analysis, ranging from basic structural studies and immune localization of selected antigens to high resolution and three-dimensional analyses for a wide array of specimens. Addition of focused ion beam scanning electron microscopy (FIB-SEM), light microscopy (LM), digital spatial profiling system and expansion of the cryo-EM technologies both at the Bethesda and RML campuses provide greater access for DIR scientists to advanced imaging technologies and spatial genomic studies. The facility provides sample preparation, imaging, and analysis, ranging from basic structural studies and immune-localization of selected antigens to high resolution and three-dimensional analyses for a wide array of specimens. EM Unit has put much effort into the critical first step of specimen preparation as evidenced by the development of application-specific chemical and cryogenic preparative techniques to optimally preserve structures of interest for visualization and interpretation. These techniques in combination with new advanced imaging technologies and the ability to plunge freeze for near native state preservation, now allows visualization of macromolecular complexes, viruses, bacteria, and host-pathogen interactions at high resolution and in three dimensions. Concurrently, our bioinformatics component continues to evolve for high-resolution analyses of increasingly complex structures, and more efficient and comprehensive data collection for higher and better throughput. In particular, the light microscopy operations now have expanded with extensive experience in studying bacteria, viruses, and human parasites and their interactions with the host, using an array of light microscopy technologies including live, low-light imaging, wide-field/deconvolution, and confocal microscopy. New personnel include interns and contract hires who bring both microscopy and data management expertise. On-going developments in both techniques and technology include but not limited to; 1) optimizing specimen preparation for immunology, 2) light microscopy (LM), 3) plunge freezing, 4) application of machine-learning, 5) light microscopy upgrades, 6) high resolution cameras and whole cell visualization though focused-ion beam scanning electron microscopy (FIB-SEM). All have allowed for an overall expansion of EM capabilities in several areas including significant improvement in 3-D visualization and resolution potential at 3 Angstroms. Simultaneously, our imaging-informatics component continually evolves with high -resolution analyses of increasingly complex structures, the ability to quantify various types of data, along with more efficient and comprehensive data collection for higher resolution and throughput. The Spatial Technologies Resource brings added capabilities by way of Nanostring, GeoMx DSP, 10X Genomics (Visium HD, and Xenium) platforms. Flow Cytometry The Flow Cytometry Section (FCS) provides the DIR with application-specific flow cytometric technologies, including instrumentation for high speed cell sorting and multi-color analysis. In addition, the FCS provides training, consultation, method development, and analysis for experiments involving flow cytometry. This mission is accomplished through the efforts of dedicated staff with >200 years of cumulated flow cytometry experience. FCS also provides cost- and time-efficient methods of obtaining fluorescent-labeled proteins or antibodies. Resources available include custom hybridoma expansion; antibody purification from ascites, antisera, and hybridoma supernatant; and coupling of purified antibodies and proteins to various fluorochromes. Major goals are to provide access to state-of-the art technologies, to help design and run experiments, to facilitate data interpretation and to provide results that are of consistent high quality. Project-specific research applications are developed for flow cytometry analysis and sorting in BSL-2, BSL-3 and BSL-4 environments. RML Genomics Research The Genomic Research Section (GRS) provides state-of-the-art next-generation sequencing, and other massively parallel genomic technologies to intramural investigators. In addition to managing high-end instrumentation, computers, and protocols involved in genome-wide assays, the GRS offers statistical analysis and bioinformatics for data interpretation. This support involves close collaboration with the investigators in both experimental design and data analysis to best utilize the power of these applications, as well as to convey a better understanding of the technology. GRS develops project-specific research applications for many analysis platforms at the DNA, RNA and protein level. The Section maintains instrumentation and resources for sequencing applications using the Illumina high throughput sequencing (NGS), including RNA-seq, DNA-seq, ChIP-seq, ATAC-seq, Rep-seq, and single-cell RNA profiling and many others. Proteins & Chemistry The Proteins & Chemistry Section develops project specific applications in the fields of proteomics, lipidomics, metabolomics, purifications, assay development, and mass spectrometry-based applications. The RTB designs qualitative and quantitative physical-chemical and biochemical methods of detection and analysis that are customized to meet the specific research needs of DIR investigators. Another major activity of the RTB involves the development of more efficient and effective methods of sample preparation with a strong orientation towards downstream mass spectrometry-based multiomics work. The Branch collaborates with DIR investigators on all facets of project design including experimental design, separation of biomolecules, sample preparation, data collection and analysis/interpretation of data. The metabolomics/lipidomics team (MetLip) is based our of RML while supporting investigators from all campuses and ICs. They use semi-targeted and targeted approaches to provide insight into biomarkers and mechanisms of disease. Visual and Medical Arts The Visual and Medical Arts Unit (VMA) at Rocky Mountain Laboratories (RML) provides highly specialized and advanced technologies to support the Division of Intramural Research (DIR) program. The unit offers a wide variety of digital technologies, involving a comprehensive suite of illustration, design services, photography, 3D animation, video, and new media to document scientific findings. In addition, the VMA provided scientific visualizations to the Office of the Director, NIAID for presentation to scientists, the public as well as US lawmakers. This unit works in close collaboration with the research community.