A central problem in ovarian cancer is late diagnosis, when the 5-year survival rate plummets to less than 50%. Ovarian cancer symptoms are vague and nonspecific, and screening is not generally effective. Because ovarian cancer is so deadly, prophylactic bilateral salpingo-oophorectomy (BSO) is often recommended for women at high risk, however BSO has fertility and health consequences. Also, for a large number of women at elevated but not extremely high risk, the best prevention action - at the best time - is unclear. It is now believed that deadly ovarian cancer may actually start in the fallopian tubes (FTs), and that precancerous cells might be detectable before they spread to the ovary. The FTs can be examined by passing a thin falloposcope through a hysteroscope and the ostia of the FT. Our long-term goal is to create an effective, minimal-invasive, and clinically reliable method to assess a woman’s FT health including detecting the earliest pre-cancerous abnormalities, to guide a physician’s and patient’s choice of appropriate prevention or treatment methods. Detecting small premalignant lesions is challenging, and we will employ three complimentary techniques with promise for this task. The first is multispectral fluorescence imaging (MFI), which has previously been shown to differentiate normal from cancerous tissue in the ovaries and FT. This instantaneous, intact tissue modality will be combined with two ex vivo cellular analysis techniques known to be sensitive to subtle pre- cancerous changes. First, Reverse Phase Protein Microarray (RPPA) measures functional cellular proteins in microscopic quantities of cells, including a number of key signaling and biochemical events that are known mediators of cancerous transformation. Second, karyometry, or computer-aided chromatin pattern analysis, has been shown in several organs to correlate with presence of early disease and risk of progression to invasive cancer. It has also shown normalization of ovary and FT tissue after chemoprevention, and has revealed the presence of a strong field effect in many organs, or the ability to sense a cancerous lesion several centimeters away from the cell collection site. The overall goal of this project is to show proof of principle of a combined method for differentiating FT samples from normal risk, high risk, and ovarian cancer patients, and to develop a method that can be used to collecting adequate FT cells for analysis. The specific aims are: Aim 1. Examine tissue-banked and newly-collected histological sections of FTs with fluorescence imaging, RPPA, and karyometry to develop an integrated algorithm that stratifies patients based on presence of disease and as a function of known patient risk status. Examine the algorithm as a function of spatial location in the FT. Aim 2: Develop a falloposcope, containing an imaging channel for navigation and MFI capability, as well as a cell sampling channel. Examine aspiration and wire scrape options for their ability to collect adequate cells for RPPA and karyometry analysis.