We are requesting funds to purchase an image analysis facility including a combination light and fluorescence microscope, multitasking computer, image analysis hardware and software. Ten separate projects requiring image analysis methodologies are identified, eight of which are currently supported by NIH. The major user group is compose of faculty members in the Departments of Anatomy, Physiology, Pharmacology and Microbiology at the West Virginia University Health Sciences Center, 12 of whom are currently funded NIH investigators. The requested image analysis instrumentation will be used by each of the group members to accomplish goals specified in or related to ongoing NIH-sponsored research projects or programs. The primary purpose of the image analysis facility will be to collect quantitative experimental data from cells or tissues viewed either directly through the light microscope using bright field, phase, differential interference, dark field, or fluorescence illumination, or indirectly by using photographs including both light and electron micrographs. The imaging needs of the group include threshholding and segmentation, image capture, size sorting, edge detection, densitometry, morphometry, low level fluorescence detection, fluorescence ratio imaging and three dimensional reconstruction and neuron analysis from tissue sections, whole mounted specimens, or micrographs. Specifically, the following list of project goals are extracted from ongoing, pending, planned NIH-funded projects that would directly benefit from the requested equipment: 1) The structural basis of neural regulation in the lung will be examined by defining the connections and distribution of chemically defined neurons in the airways. 2) Seasonal changes in hypothalamic-neuroendocrine interactions related to seasonal reproductive cycles will be studied. 3) The effects of diet on the process of myelination will be examined. 4) Autoradiography analysis of cell membrane receptors will be used to study the process of morphine tolerance in gastrointestinal neurons. 5) Pharmacodynamic mechanisms of glomerular contractility in pregnancy and aging will be evaluated. 6) Changes in mRNA and protein levels in human breast cancer cells will be studied using in situ hybridization and immunocytochemical methodologies. 7) Morphometric analysis of changes in thyroid tissue as related to thyroid blood flow will be examined. 8) The somatotopic organization of spinal afferent projections will be evaluated using neural tracers. 9) Different swim forms of different spirochetes will be evaluated. 10) Calcium shifts in cultured astrocytes will be measured. The requested image analysis instrument will be housed in a dedicated space separated from but adjacent to the laboratory of the principal investigator enabling the collaborative arrangements already established between the project investigators and the PI to continue. A state-of-the-art image analysis facility will enhance the research programs of the user group by providing accurate, reproducible and efficient acquisition of quantitative data about structural and molecular aspects of biological systems.
