Laser capture microdissection has become a primary research technique to isolate specific cell populations from tissues or cell culture affording further sub-cellular techniques to be conducted on highly purified cells rather than a heterogeneous tissue. This proposal requests support for the purchase of a Leica LMD7000 laser capture microscope, to be maintained in an institutional Shared Resource, the Molecular Pathology Core. Access to this instrumentation is crucial to the work of two major user groups, namely, Center for Immunology and Transplantation and Laboratory of Inflammation Biology and Surgical Sciences, at the University of Florida. Administration, maintenance, and high-level technical assistance and training will be provided by experienced staff in the Molecular Pathology Core. All user training and biosafety issues have been addressed. Strong institutional commitment is demonstrated by the support of College of Medicine Dean, Dr. Michael Good, and Senior Associate Dean, Dr. Steven Sugrue, and Department of Pathology, Immunology and Laboratory Medicine Chairman, Dr. Michael Clare-Salzler. The Leica LMD7000 was selected as the most suitable for a multi-user facility due to its greater flexibility for a wide range of sampe types and clearly fulfills an unmet need as the institution does not have this instrumentation available through a shared resource. The Leica instrument has several ideal features including intuitive and simple software to automate sample dissection, 4-tube extraction capabilities, brightfield and fluorescence, cell culture, and superior objectives for sharp laser cutting. The major users highlighted in this proposal are well-funded by the NIH with active and pending grants requiring this instrumentation. Each would benefit greatly from having institutional access to a laser capture instrument. Martha Campbell-Thompson and Mark Atkinson will use the LMD7000 to study infiltrating lymphocytes and islets to understand mechanisms of beta cell destruction and islet regeneration for type 1 diabetes. Clayton Mathews will conduct research on the basic biology of mitochondria in beta cells and altered glucose metabolism through genomic analyses of isolated islets. Laurence Morel studies various immune cell populations in a model of systemic lupus and is currently subcontracting the laser capture studies to a collaborator at another institution. Lyle Moldawer studies host responses to acute inflammatory processes in sepsis and systemic inflammatory response syndromes and to chronic inflammatory processes, including cancer and AIDS-associated cachexia. He too has active NIH funding for studies with laser capture microdissection for arterial venous fistula. Finally, the instrument will be availabl to other investigators through this shared resource using a fee-for-service mechanism for training and assisted or unassisted use.
|Zhu, Ying; Piehowski, Paul D; Zhao, Rui et al. (2018) Nanodroplet processing platform for deep and quantitative proteome profiling of 10-100 mammalian cells. Nat Commun 9:882|