The major goal of this grant is the understanding of slit-scan light scatter signals and the information they provide. This will be accomplished through the following tasks: (1) The completion of the addition of low angle forward and large angle light scatter sensors to two multidimensional slit-scan flow systems. (2) The reassembly of a Correlation System to provide two orthogonal fluorescence images of cells in flow together with fluorescence and light scatter slit-scan contours. (3) The collection and correlation of slit-scan light scatter and fluorescence contours. Because the two contours are perfectly correlated, this will permit identification in the slit-scan light scatter contour of the exact point at which the cytoplasm and nuclear boundaries of the cell intersect the slit of laser illumination. Specific cell types will be analyzed to aid in the understanding of the information available from the slit-scan light scatter contours. Of particular interest are nuclear and cell boundaries together with information on intracellular structure. (4) The investigation of detector geometry to optimize morphological boundary, clump, internal structure, and orientation information from slit-scan light scatter contours on cells in flow. (5) The collection and correlation of slit-scan light scatter contours with slit-scan fluorescence contours and two orthogonal fluorescence images of cells in flow. This data will aid in the study and understanding of the effects of cell orientation in flow on slit-scan light scatter signals and the use of slit-scan light scatter signals to provide information on cell orientation in flow, internal structure, multinucleation, and clumping of cells, and the identification of low resolution morphological features. (6) Increasing the resolution on one system to two microns to permit the study and understanding of slit-scan light scatter information on other cell types of interest. (7) Parallel theoretical studies and guidance to aid in hardware design, study direction, and the understanding of slit-scan light scatter information. This study is new and unique and will provide information relevant to cancer detection utilizing multidimensional slit-scan instrumentation, as well as be useful in conventional flow cytometry.
