Duke University Medical Center currently has 21 investigators in need of state-of-the-art technology that combines multispectral (brightfield, darkfield, and fluorescence) imaging with flow cytometry. These investigators come from a wide-range of departments and disciplines (Biochemistry, Cell Biology, Immunology, Medicine, Microbiology and Genetics, Pediatrics, Radiation Oncology, Pharmacology and Cancer Biology). In addition, these investigators have a diverse array of research interests from cancer biology, cell biology, cellular signaling, immunobiology, neurobiology, receptor trafficking, stem cell biology and vascular biology. The currently funded NIH projects of each of these investigators would benefit tremendously from the ImageStream System available from Amnis Corporation. The ImageStream multispectral imaging flow cytometer produces high resolution brightfield, darkfield, and fluorescence images of cells prepared in suspension at rates exceeding 100 cells per second. The IDEAS"""""""" analysis software quantifies over 500 morphometric and photometric parameters for each cell based on its imagery, including parameters that measure sub-cellular location of probes. By combining the statistical power afforded by large sample sizes common to flow cytometry with numerical quantification of the high information content present in microscopic images, this technology enables many novel cell image-based applications, including but not limited to analysis of apoptosis, autophagy, chemotaxis, immune synapse formation, receptor internalization and trafficking and signal transduction (cell membrane localization, nuclear translocation). Importantly, many of these factors can be assessed simultaneously, allowing real time assessment of the effects of these parameters on each other (i.e. receptor trafficking on signaling outcomes). In addition, this technology also allows assessment of subtle and/or rare events that would not be captured by whole sample analysis of these same parameters. As such, use of the ImageStream System will have tremendous impact on a broad diversity of NIH funded research projects, not only facilitating their completion, but enhancing the goals of these projects and allowing researchers at Duke University to more definitively answer key questions of direct relevance to human health, including cancer, heart disease and immune system, pulmonary and vascular disorders.