In the last year, 40 researchers have used the resources of the Laboratory of Cellular and Molecular Biology Microscopy Core. While most of the researchers come from the Laboratory of Cellular and Molecular Biology, the Core has been used by scientists from the Laboratory of Immune Cell Biology,the Cell and Cancer Biology Branch and The University of Maryland Department of Physics. Almost all of the Principal Investigators in the Laboratory of Cellular and Molecular Biology have projects that involve the Core facility. Dr. Lawrence Samelson uses Core resources for the project """"""""Biochemical Basis of T Cell Activation"""""""". Researchers working with Dr. Carol Clayberger have used the facility for the projects """"""""Regulation of RANTES Expression in T Lymphocytes"""""""" and """"""""Function of Granulysin"""""""". The Core has been involved with the project """"""""Cbl Proteins as Regulators of Tyrosine Kinase Signaling""""""""from Dr. Stanley Lipkowitz. Dr. Carole Parent's projects, """"""""Signaling Events Regulating Chemotaxis"""""""" and""""""""Chemotactic Signals Reulating Human Neutrophil and Breast Metastatic Migration"""""""" use Core instruments. Dr. Paul Randazzo has made extensive use of the Core for the projects """"""""Regulation of focal adhesions"""""""" and """"""""Turnover of invadopodia"""""""". The Core facility has assisted Dr.Jeffry Rubin with the project """"""""Casein Kinase 1 Delta in Wnt Signaling and Beyond"""""""". Dr. Ying Zhang uses Core instruments for the project """"""""Molecular Mechanisms of TGF-beta Signaling Pathway"""""""". In addition, the Core facility has been used by personnel working with Principal Investigators from other groups including work with Dr. Michael Bustin on the nucleosome binding protein HMGN1 and the role of heterochromatin formation in cell migration. This research usually involves the use of a Zeiss 510 Laser Scanning Confocal Microscope or a PerkinElmer UltraView Spinning Disk Confocal Microscope, however we are seeing increased use of our Total Internal Reflection Fluorescence (TIRF) microscope. Most of the users view immunofluorescent staining on fixed samples with the Zeiss 501 LSCM while the spinning disk confocal and TIRF microscopes are generally used for live cell imaging. We routinely use our TIRF microscope for PhotoActivation Localization Microscopy (PALM). This high resolution technique has allowed us to determine the location of single proteins clustered in signaling complexes in T cells with an accuracy of around 20 nm, well below the diffraction limit of visible light. The TIRF system is also being used for the projects """"""""Regulation of focal adhesions"""""""" and """"""""Turnover of invadopodia"""""""" with Dr. Paul Randazzo.
| Guittard, Geoffrey; Dios-Esponera, Ana; Palmer, Douglas C et al. (2018) The Cish SH2 domain is essential for PLC-?1 regulation in TCR stimulated CD8+ T cells. Sci Rep 8:5336 |
| Dutta, Debjani; Barr, Valarie A; Akpan, Itoro et al. (2017) Recruitment of calcineurin to the TCR positively regulates T cell activation. Nat Immunol 18:196-204 |
| Barr, Valarie A; Yi, Jason; Samelson, Lawrence E (2017) Super-resolution Analysis of TCR-Dependent Signaling: Single-Molecule Localization Microscopy. Methods Mol Biol 1584:183-206 |
| Yi, Jason; Manna, Asit; Barr, Valarie A et al. (2017) Highly Multiplexed, Super-resolution Imaging of T Cells Using madSTORM. J Vis Exp : |
| Yi, Jason; Manna, Asit; Barr, Valarie A et al. (2016) madSTORM: a superresolution technique for large-scale multiplexing at single-molecule accuracy. Mol Biol Cell 27:3591-3600 |
| Sherman, Eilon; Barr, Valarie A; Merrill, Robert K et al. (2016) Hierarchical nanostructure and synergy of multimolecular signalling complexes. Nat Commun 7:12161 |
| Barr, Valarie A; Sherman, Eilon; Yi, Jason et al. (2016) Development of nanoscale structure in LAT-based signaling complexes. J Cell Sci 129:4548-4562 |
| Balagopalan, Lakshmi; Kortum, Robert L; Coussens, Nathan P et al. (2015) The linker for activation of T cells (LAT) signaling hub: from signaling complexes to microclusters. J Biol Chem 290:26422-9 |
| Zvezdova, Ekaterina; Lee, Jan; El-Khoury, Dalal et al. (2014) In vivo functional mapping of the conserved protein domains within murine Themis1. Immunol Cell Biol 92:721-8 |
| Sherman, Eilon; Barr, Valarie A; Samelson, Lawrence E (2013) Resolving multi-molecular protein interactions by photoactivated localization microscopy. Methods 59:261-9 |
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