The objective of this project is to provide researchers with advanced graphical and numerical methods for fully utilizing data obtained from the optical sectioning of cells and cell-nuclei. The principal application will be in the study of chromosome arrangement in interphase cells. Recent advances in the development of fluorescent nuclei acid probes for chromosome-specific repetitive sequences and collections of unique sequences (composite probes) have permitted the detection of individual chromosomes in the interphase nucleus. Through the techniques of optical sectioning and 3-dimensional image reconstruction, it is now possible to study the spatial distribution of individual chromosomes within the cell nucleus and its possible involvement in the origin of translocations and aneuploidies. This will involve measuring the location of each chromosome, relative to the center and surface of the nucleus, and its proximity to its homolog and other chromosomes within the nucleus. This information will be correlated for data from many cells, to establish the natural variability in such data. This will permit a study of the role of nuclear organization on differentiation and transformation. To accomplish these objectives, we will: 1) Develop methods for identifying and displaying objects in complex 3-dimensional images. Objects will be detected using 3D edge finding techniques and displayed using queuing and/or stereo displays. 2) Develop quantitative methods for extracting features of 3-dimensional objects. Features of interest include shape, volume, surface area, integrated fluorescence intensity, location within the cell or cell nucleus, and distance to other objects within the cell/nucleus. 3) Develop statistical to determine the 3-dimensional regularity of organization of objects within cells and cell nuclei. 4) Develop a data base to archive the raw data from 3-dimensional images, parameters used for rendering and display, and results of analysis.
