Shared Instrumentation: Amnis ImageStream System

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DESCRIPTION (provided by applicant): Funding is requested for the purchase of an Amnis ImageStream multispectral imaging flow cytometer which can perform high-throughput analysis of cell populations and cellular processes. This instrument essentially combines the visual power of microscopy, the quantitative ability of image analysis, and the statistical rigor of flow cytometry in a single platform to create exceptional new experimental capabilities. Although destined to become the single analysis platform of choice for cell analysis, this technology has only recently become available such that there are currently still relatively few of these instruments in research institutions. Compared to traditional flow cytometry, the ImageStream also uses data derived from cell morphology which provides a more comprehensive analysis of the sample, while still providing all of the fluorescent information. The feature that is truly enabling, however, is the added ability to identify the intracellular location of the fluorescence signal through cell morphological analysis. The ImageStream also makes it possible to track signaling events or protein movement in a spatial/temporal manner, within or between cells. To determine whether rare events in the data are real or due to artifacts, individual specific signals can be visualized within a specific image using the ImageStream. This also holds true for accuracy in gating strategies. With the introduction of Extended Depth of Field (EDF) technology, the ImageStream can also generate confocal-like images. This technology really enhances the ability to look at FISH spots and co-localization events. As a result of these technological advances, the ImageStream can perform an extensive range of applications including: nuclear translocation, internalization molecular trafficking, molecular co-localization, morphological classification, nuclear foci spot counting and apoptosis detection. In this proposal we present 13 individual NIH funded grants that would directly benefit from access to this novel technology. Specifically the instrument will enhance projects in genomics studies through the application of FISH and detection of phosphorylation. Work in the fields of cellular immunology will benefit considerably from the capacity of the instrument to detect rare cellular subpopulations and cancer research projects which are dependent on the ability to perform co-localization, nuclear translocation studies, apoptosis, intercellular localization and morphological classification will similarly be enhanced. The ImageStream will be incorporated into the Flow cytmoetry core facility in the MCG Cancer Center and its day to day operation will be overseen by the Flow core facility manager. The PI is the director of the MCGCC core facilities for clinical and translational research who will oversee its operation, act as chair of the advisory committee and resolve issues arising with the MCGCC administration.