In order to determine the biological origin of the optical signatures of intracellular and extracellular constituents we propose to further develop and study an in-vitro tissue slice model.
Biopsies will be obtained from women undergoing oophorectomy.. Embedded tissue samples will be sectioned with a precision cut tissue slicer, incubated, and microscopically examined. We will determine the duration of the tissue slice viability. Development of the ovarian tissue slice model will be in collaboration with investigators from the University of Arizona, College of Pharmacy, and will have application in many future studies.
We will explore the optical signature of tissue constituents, quantify them and mapping their spatial distribution within the tissue slice with microscopy techniques. Images will be obtained with white light and two-photon excitation microscopy. White light reflectance images will provide scattering information while fluorescence images will determine spatial distribution of fluorophores. Optical properties from structures identified in these images will be correlated with histopathologic analysis. The focus will be to identify structures contributing to the contrast between normal and neoplastic transformed ovarian tissue. Imaging will be performed at the University of Arizona in the user facilities of the Arizona Research Laboratories.
Our efforts to achieve aim 1 and 2 require data from biopsy samples from pre and postmenopausal women because hormonal status will affect tissue composition. Our samples will be studied ex-vivo because our in-vitro tissue model will keep tissue alive during microscopy studies.

Figure: Single photon and two photon images of the ovarian surface epithelium as well as the stroma. Left: 2P excitation at 720 nm and fluorescence emission recorded after IR filtering. Strongest signal is originating from the epithelial tissue layer. Structural contrast can also be observed in the stroma. Right: The same site excited at 458 nm in single photon mode shows stronger contribution from the epithelium and less structural contrast. Emission at 458nm excitation is consistent with FAD and emission from 2P excitation at 720nm with NAD(P)H and structural proteins. A bar is 200 mm.