CMB Research Support Specialists provided technical expertise and support to NIAID Principal Investigators, assisting them with true cage to benchside support. Both pathology and technical proficiency resulted in numerous co-authorship opportunities. The CMB has successfully maintained a gnotobiotic breeding and study facility, which consists of bio-exclusion units designed to keep the mice from becoming colonized with any adventitious microorganisms. Germ-free mice are free of all aerobic and anaerobic organisms with the possible exception of endogenous viruses. Breeding colonies and mice on study are maintained in isolators provided with HEPA-filtered air and autoclaved food, bedding, and supplies. Strict SOPs are followed to maintain the mice in a germ-free state. The Mouse Genetics and Gene Modification (MGGM) section was established as a new section at CMB/NIAID to provide highly sophisticated services related to gene modification in mice and long term storage of transgenic and gene knockout mouse lines. MGGM provided these services to over 25 different labs at the NIAID. Recently, MGGM has established a microinjection set up for the production of transgenic animals by microinjection of DNA and also for microinjection of CRISPR/cas gRNA for genome editing in mouse embryos. Similarly, a secondary set up is being established at to provide MNV and helicobacter free animals to the NIAID investigators. MGGM is also setting up a tissue culture lab for ES cell technology for gene knock in/knockout projects via homologous recombination as well as by CRISPR/cas system. This year, the MGGM provided sperm cryopreservation (47 completed or in progress), embryo cryopreservation (7 completed or in progress), mouse line rederivation (11), microinjection of embryos (1) and ovarian transfers (2). The Infectious Disease Pathogenesis Section (IDPS) aids investigators in animal study design, necropsy assistance and guidance, tissue sample collection protocols, education on the anatomy, physiology, and pathology terminology related to a specific animal model, in efforts to prepare investigators for national and international presentations. We have significantly improved the quality of IDPS pathology support given to NIAID investigators. The IDPS has made a number of functional changes and improvements to the services we provide to the greater NIAID community of investigators. We are currently in the process of moving the IDPS to a full-service histopathology lab with our recent acquisition of a Hematoxylin and Eosin (H&E) stainer and our upcoming acquisition of a fully-automated tissue processor and embedding station. We will also acquire an automated machine that can process specimen for both immunohistochemistry (IHC) and In-situ Hybridization (ISH). In the not too distant future, the presence of these technologies will give us the ability to process certain tissues and perform these different analyses within our lab space. This tissue processing ability will allow for more options and quicker turn-around for the investigator. In addition to staining, we have also identified a digital pathology system that will help us scan histology slides at high-resolution and work with investigators in a virtual setting and allow a certain degree of additional methods of data analysis. This year, we have generated a number of pathology data sets for investigators for which publications are currently in progress, have been submitted for publication or, in particular, ones that have been accepted in highly respected peer-reviewed journals such as Nature. Our main objective will be to continuing to grow, direct the IDPS into the future with cutting-edge technology, make the IDPS a self-sufficient, full service histology laboratory, and to push the envelope as it relates to molecular pathology support for the greater intramural research team within the National Institutes of Allergy and Infectious Disease.
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