This request is for an Arcturus AutoPix II, which is an automated laser capture microdissection microscope equipped for both brightfield and fluorescent operation. The system will significantly enhance the research of 4 major users who are conducting NIH-funded R01 research projects and 2 minor users, one of whom holds an R21. Drs. Wyrick and Raulston would use LCM technology to analyze tissue subpopulations of genital epithelial cells that are susceptible to infection by Chlamydia trachomatis. Such samples would be examined for transcription and/or protein expression of (i) estrogen receptors involved in Chlamydia attachment, (ii) chemokines produced by infected cells, and (iii) Chlamydial iron-responsive genes and proteins; uninfected, non-susceptible genital cells within the same tissue would serve as comparative controls. Dr. Hoover will use the instrument to isolate contents of single or multiple neurons of guinea pig intracardiac ganglia and use RT PCR to determine the presence of mRNA coding for tachykinin receptors and receptors for calcitonin gene-related peptide and related mediators. Dr. Li will employ this equipment to examine the role of Toll-like receptors (TLRs) in myocardial ischemia/reperfusion injury. In ischemic and non-ischemic tissue captured through the microdissection, protein expression and transcription for TLRs, selected cytokines and for various protein kinases will be assayed. The Arcturus instrument is the standard instrument of its kind. The East Tennessee State University College of Medicine has a strong institutional commitment to the acquisition and maintenance of this microscope system. A highly experienced and full time research assistant, Johanna Preiszner, Ph.D., will oversee the day to day operation of the microscope. A room of about 100 square feet in the Department of Pathology will house the system. The College of Medicine has also agreed to underwrite the service contract for the instrument after the initial warranty year. ? ?
