The investigators will focus their research on fully developing the automated instrumentation capabilities of microscopic systems that offer the potential for quantifying the 3-D shape parameters, and changes in shape parameters over time, of microscopic-scale biological specimens. They plan to continue their program of studying diversity of complementary imaging modalities, including confocal microscopy, stereo microscopy, and conventional optical sectioning microscopy. The program of study will encompass microscopic data acquisition via various methods. The images obtained will be subjected to appropriate filtering schemes base on the physical principles of image formation to sharpen and improve image clarity in individual micrographs and to remove information spurious to that of the local specimen structure. The filtering techniques, which will require considerable analytic study in their development, will be specific to each optical system used. Finally, the aggregate local optical data will be assembled and analyzed via sophisticated numerical instrumentation techniques to reconstruct the 3-D structure of a given imaged specimen and to quantify its size and shape characteristics as it evolves over time. The methods developed will be implemented and evaluated on a broad range of specimen types, including stained and unstained single and multiple celled subjects. Several different tissue preparations will be used to study the time evolution of specimen morphology over time during various biological processes.
