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)
Project #
Application #
Study Section
Special Emphasis Panel (ZAR1-KM)
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Yale University
New Haven
United States
Zip Code
Shulman, Ziv; Gitlin, Alexander D; Weinstein, Jason S et al. (2014) Dynamic signaling by T follicular helper cells during germinal center B cell selection. Science 345:1058-62
Bockenstedt, Linda K; Gonzalez, David; Mao, Jialing et al. (2014) What ticks do under your skin: two-photon intravital imaging of Ixodes scapularis feeding in the presence of the lyme disease spirochete. Yale J Biol Med 87:3-13
Weinstein, Jason S; Bertino, Sarah A; Hernandez, Sairy G et al. (2014) B cells in T follicular helper cell development and function: separable roles in delivery of ICOS ligand and antigen. J Immunol 192:3166-79
Deschene, Elizabeth R; Myung, Peggy; Rompolas, Panteleimon et al. (2014) ?-Catenin activation regulates tissue growth non-cell autonomously in the hair stem cell niche. Science 343:1353-6
Ray, John P; Marshall, Heather D; Laidlaw, Brian J et al. (2014) Transcription factor STAT3 and type I interferons are corepressive insulators for differentiation of follicular helper and T helper 1 cells. Immunity 40:367-77
Teichmann, Lino L; Schenten, Dominik; Medzhitov, Ruslan et al. (2013) Signals via the adaptor MyD88 in B cells and DCs make distinct and synergistic contributions to immune activation and tissue damage in lupus. Immunity 38:528-40
Kumamoto, Yosuke; Linehan, Melissa; Weinstein, Jason S et al. (2013) CD301býýý dermal dendritic cells drive T helper 2 cell-mediated immunity. Immunity 39:733-43
Truman, Lucy A; A-Gonzalez, Noelia; Bentley, Kevin L et al. (2013) Lymphatic vessel function in head and neck inflammation. Lymphat Res Biol 11:187-92
Bertino, Sarah A; Craft, Joe (2013) Roquin paralogs add a new dimension to ICOS regulation. Immunity 38:624-6
Nickerson, Kevin M; Cullen, Jaime L; Kashgarian, Michael et al. (2013) Exacerbated autoimmunity in the absence of TLR9 in MRL.Fas(lpr) mice depends on Ifnar1. J Immunol 190:3889-94

Showing the most recent 10 out of 29 publications