Two-photon laser scanning microscopy is a technique that can be applied to the imaging of fluorescently tagged cells within intact tissue using point scanning microscopes and pulsed infra-red lasers. This approach can be employed with either excised bulk tissue or with live, anesthetized animals, a technique referred to as intravital microscopy (IVM). In order to perform this technique, equipment is required that is prohibitively expensive to all but a few investigators. Yale University School of Medicine investigators are fortunate in that the school has already invested in a two-photon laser and custom designed laser scanning microscope equipped with sensitive external detectors that allow for the collection of low level light. A unique microscopy suite for use by this Core C permits the preparation of animals and their subsequent imaging to occur in adjacent rooms. This facility has been subsidized through a P30 Core grant and made available to all members of the YRDRCC. For a multitude of these investigators, the application of this techniques to their research has greatly accelerated the progress of their research and provided substantial insights into their in vivo biological questions in a way that is not possible with traditional in vitro assays. Together with the Core PI, the facility operator has consulted with investigators to ensure that experimentation came to fruition quickly and efficiently. P30 funding has made possible substantial methods development that enabled many novel applications, and expanded the fluorophore combinations and tissue types that could be imaged. Nevertheless, without additional P30 Core funding, many barriers that prevent investigators from applying this approach to their in vivo research would resurface. For example, the cost of the service contracts for maintenance of the laser and microscope would require that facility user fees be greatly increased, posing a significant financial deterrent that would prohibit its use by many investigators, especially junior PIs. The additional skills needed for the animal anesthesia, surgery and restraint during IVM appear daunting to most researchers, despite the training and advice that we have made available. Therefore, this application proposes to expand the capabilities and further promote the In Vivo Imaging Core Facility. P30 funding would not only reduce these barriers but also acquire additional methodologies to query as yet inaccessible biological events. The proposed IVICF would have a dramatic impact on the pace of research into cell-interaction, differentiation and migration in vivo in both health and in rheumatic diseases.

Public Health Relevance

This application proposes to expand the capabilities and further promote an In Vivo Imaging Core Facility that allows fluorescently tagged cells to be imaged within living anesthetized mice. P30 funding would reduce numerous significant barriers and allow for the development of additional methodologies. The proposed core would have a dramatic impact on the pace of research in both health and in rheumatic diseases.

National Institute of Health (NIH)
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Center Core Grants (P30)
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Special Emphasis Panel (ZAR1-KM (M1))
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Yale University
New Haven
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Weinstein, Jason S; Laidlaw, Brian J; Lu, Yisi et al. (2018) STAT4 and T-bet control follicular helper T cell development in viral infections. J Exp Med 215:337-355
Fistonich, Chris; Zehentmeier, Sandra; Bednarski, Jeffrey J et al. (2018) Cell circuits between B cell progenitors and IL-7+ mesenchymal progenitor cells control B cell development. J Exp Med 215:2586-2599
Gies, Vincent; Schickel, Jean-Nicolas; Jung, Sophie et al. (2018) Impaired TLR9 responses in B cells from patients with systemic lupus erythematosus. JCI Insight 3:
Gonzalez, David G; Cote, Christine M; Patel, Jaymin R et al. (2018) Nonredundant Roles of IL-21 and IL-4 in the Phased Initiation of Germinal Center B Cells and Subsequent Self-Renewal Transitions. J Immunol 201:3569-3579
Manfredo Vieira, S; Hiltensperger, M; Kumar, V et al. (2018) Translocation of a gut pathobiont drives autoimmunity in mice and humans. Science 359:1156-1161
Greiling, Teri M; Dehner, Carina; Chen, Xinguo et al. (2018) Commensal orthologs of the human autoantigen Ro60 as triggers of autoimmunity in lupus. Sci Transl Med 10:
Kim, Sang Taek; Choi, Jin-Young; Lainez, Begona et al. (2018) Human Extrafollicular CD4+ Th Cells Help Memory B Cells Produce Igs. J Immunol 201:1359-1372
Ip, W K Eddie; Hoshi, Namiko; Shouval, Dror S et al. (2017) Anti-inflammatory effect of IL-10 mediated by metabolic reprogramming of macrophages. Science 356:513-519
Swartz, Kelsey L; Wood, Scott N; Murthy, Tushar et al. (2017) E2F-2 Promotes Nuclear Condensation and Enucleation of Terminally Differentiated Erythroblasts. Mol Cell Biol 37:
Di Pietro, Caterina; Zhang, Ping-Xia; O'Rourke, Timothy K et al. (2017) Ezrin links CFTR to TLR4 signaling to orchestrate anti-bacterial immune response in macrophages. Sci Rep 7:10882

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