? Two-photon microscopy allows for the visualization of cellular processes, such as cell? migration, cell-to-cell and cell-to-matrix interactions, subcellular organelle function? (mitochondria), cellular signaling (Ca2+, reactive oxygen species, and phospholipid? uptake), characterization of tumor growth, drug targeting, and protein expression within? the three-dimensional, natural environment of intact organs in living animals. During the? past few years, two-photon microscopy has revolutionized our view of developmental,? homeostatic, and pathologic processes. We propose to establish an intravital two-photon? microscopy system at The Institute for Environmental Medicine (IFEM) through the? acquisition of a Leica TCS SP5 Spectral Confocal and Two-Photon Microscope in? combination with a Spectra-Physics Broadband Mai-Tai femtosecond-pulsed Laser. The? major users of the proposed instrument will be a group of School of Medicine? Investigators drawn from a wide range of disciplines, including Physiology, Pathology,? Emergency Medicine, Pulmonary Medicine, Cancer Biology, Rheumatology,? Hematology/Oncology and Immunology research. The need to translate experimental? data from culture models into living animals, and to expand static information of dynamic? cellular processes into four- dimensional kinetic analyses in vivo has greatly augmented? the need for intra-vital imaging at the IFEM. Such research cannot be conducted by? equipment currently available. Promising preliminary studies documenting ROS? production in the lung, vascular targeting of nanocarriers, trafficking of T-cell progenitors? in vivo, and immune cell interactions during inflammatory and infectious diseases has? demonstrated the feasibility of two-photon microscopy for the proposed projects. The? instrument will be centrally located in the School of Medicine to enable convenient? access for all users, will be maintained by the IFEM under the supervision of the PI and? Microscope Advisory Committee. Investigators from diverse fields will have new? opportunities for interaction and the initiation of interdisciplinary research projects, which? will significantly advance their NIH-funded projects. It is anticipated that the instrument? will help to obtain novel insights into the normal cellular physiology, drug delivery, and? various pathologies involved with aging, cancer, acute lung injury, transplantation,? pulmonary hypertension, atherosclerosis, autoimmunity, and chronic infections.? Relevance: The purpose of the 2-Photon microscope is to facilitate increased interaction? among clinical investigators and basic scientists who are dedicated towards the study of? various disease models and their possible treatment. This instrument will be invaluable? in translating in vitro cell culture studies to intact organ or live animal. Overall, the? equipment will enhance and expand IFEM Core to provide state of the art support to the? School of Medicine at Penn.
| Shuvaev, Vladimir V; Han, Jingyan; Yu, Kevin J et al. (2011) PECAM-targeted delivery of SOD inhibits endothelial inflammatory response. FASEB J 25:348-57 |
| Davis, Christiana W; Hawkins, Brian J; Ramasamy, Subbiah et al. (2010) Nitration of the mitochondrial complex I subunit NDUFB8 elicits RIP1- and RIP3-mediated necrosis. Free Radic Biol Med 48:306-17 |
| Hawkins, Brian J; Levin, Mark D; Doonan, Patrick J et al. (2010) Mitochondrial complex II prevents hypoxic but not calcium- and proapoptotic Bcl-2 protein-induced mitochondrial membrane potential loss. J Biol Chem 285:26494-505 |
| Madesh, Muniswamy; Zong, Wei-Xing; Hawkins, Brian J et al. (2009) Execution of superoxide-induced cell death by the proapoptotic Bcl-2-related proteins Bid and Bak. Mol Cell Biol 29:3099-112 |
