CMB Research Support Specialists provide 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 in 2015 as a new section at CMB/NIAID to provide highly sophisticated services related to gene modification in embryos and embryonic stem (ES) cells via CRISPR/cas9 technologies as well as cryopreservation of mouse lines for long term storage. MGGM has streamlined all the techniques for creating gene edited mice via CRISPR/cas9 at Bldg. 50 and Twinbrook 3. Two microinjection specialists were hired to provide gene editing and transgenic services. The Building 50 component will provide MNV and helicobacter free animals to the NIAID investigators. A tissue culture lab has been established at Twinbrook 3 for ES cell technology for gene knock in/knockout via homologous recombination as well as by CRISPR/cas9 system. During this year, MGGM provided services to over 30 individual labs as follows; 1. Over 30 CRISPR/cas9 positive animals were created for five (5) different sets of gRNAs that contain InDels in different genes including some animals with InDels at both alleles 2. MGGM has also generated transgenic animals via microinjection of large DNA constructs. These animals were generated by the double DNA injections. 3. MGGM has also completed over 50projects for sperm cryopreservation, embryo cryopreservation and rederivation of mouse lines at TB3 and Bldg. 50. Sperm and embryo cryopreservation was confirmed by QC assay by generating live born pups. The Infectious Disease Pathogenesis Section (IDPS) aids investigators NIAID investigators in the culmination and execution of a number of activities related to animal studies including, but not limited to; animal study design, necropsy instruction and assistance, tissue sample collection protocols, education on the anatomy, physiology, and the appropriate pathology terminology related to a specific animal model. The aforementioned activities are performed in effort to prepare investigators for national and international presentations and manuscript preparation and submission. We have significantly improved the quality of IDPS pathology support given to NIAID investigators and the level of communication/collaboration between the IDPS and the laboratories within NIAID. The IDPS has made a number of functional changes and improvements to the services provided to the greater NIAID community of investigators. We are currently in the process of transitioning the IDPS to a full-service histopathology lab with our recent acquisition and future implementation of a Hematoxylin and Eosin (H&E) stainer. In addition, we have recently acquired a fully-automated tissue processor, embedding station and digital slide scanner. The presence of these technologies will now provide us with the ability to process a number of tissue types and perform various analyses within our laboratory space. In-house tissue processing will allow for more options and quicker turn-around for the investigator. The digital pathology slide scanning system allows us to scan slides at high-resolution and work with investigators remotely and within a virtual setting and provides additional methods of data analysis (quantitative). This year, we have generated a number of pathology data sets for investigators for which publications that are currently in progress, have been submitted for publication or, in particular, ones that have been accepted in highly respected peer-reviewed journals including Nature, Nature Immunology, and Journal of Infectious Diseases, just to name a few. Our main objective will be to continuing to grow, direct the IDPS into the future with cutting-edge technology, and make the IDPS a self-sufficient full-service histology laboratory, and continue to raise the bar regarding molecular pathology support for the greater intramural research team within the National Institutes of Allergy and Infectious Disease.

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