Measuring whole body fluorescence and movement in freely swimming transgenic zebrafish for measuring stress from exposure to environmental stressors

Toyon Research Corporation, in collaboration with the University of California at Santa Barbara (UCSB), proposes developing a turnkey system consisting of both hardware and software necessary to measure whole-body fluorescence in freely swimming zebrafish. The proposed system will consist of a test domain and a number of imaging sensors sensitive to fluorescent illumination; it will be utilized to measure the effects of stress from environmental contaminants or stressors such as mercury, lead, or temperature. At the heart of the proposed technical solution are advanced image processing algorithms capable of detecting and tracking large numbers of dim fluorescent signatures in the presence of partial occlusions. In Phase I, Toyon proposes conducting architecture and system design; data collection of fluorescent zebrafish in facilities owned by subcontractor UCSB; development of algorithms for three-dimensional spatiotemporal tracking of zebrafish across multiple sensors; implementation of a functional graphical user interface; and analysis of illumination to minimize measured system noise. In planned Phase II development, two versions of the proposed system will be built to test and refine the architecture, algorithms, and graphical user interface developed during the Phase I efforts. Particular attention shall be applied to ensuring that measurements are repeatable and measured noise is manageable.