The objective is to develop a practical method for measurement of cochlear morphometric and physiological responses from fluorescent microscope images of cochlear tissue. Fluorescent molecular probes are chosen for the particular purpose of the experiments, i.e., marking the location and distribution of actin structural proteins in the organ of Corti. The method quantifies the spatial three dimensional extent of the fluorescent intensity from living or fixed specimens of the organ of Corti, or isolated cochlear hair cells. This is accomplished by focusing a microscope to obtain a series of images which are then operated upon by a unique computational procedure giving restored quantitative images for further analysis. The novelty of the measurement system is the incorporation of prior knowledge about the nature of the cochlear images. The phase I objective is to implement the method as a convenient procedure and analyze the F-actin distribution in an in vitro preparation of the organ of Corti. Phase I has three Specific Aims: 1) Automate the image acquisition and analysis procedure, and further develop the computational algorithm; 2) Study the image distortion introduced by measurements from the organ of Corti; and 3) Map the spatial distribution of actin in the supporting cells in the organ of Corti.
