Project Summary/Abstract This proposal is a request for funding for an Amnis ImageStreamX MarkII Imaging Flow Cytometer by Luminex Corporation. The instrument will be permanently placed within the University of Utah Flow Cytometry Core Facility. All operational oversight will be carried out by the qualified staff of the Flow Facility and its established management structures. As an analytical instrumentation core facility, we strive to provide intellectual expertise in cytometry as well as the physical tools needed by a broad range of biological disciplines to answer a widening array of inquiries. More specifically, for a cytometrist, this translates into measuring both intrinsic and extrinsic properties of cells within a heterogeneous population. Historically, microscopy has been the source of definitive fluorescent localization and morphological information, while flow cytometry has carved a niche of high throughput, high parameter, suspension cell analysis. Unfortunately, neither of these indispensible technologies has been capable of providing the full breath of data needed for a sizable set of specialized applications. These applications predominantly consist of suspension or non-adherent cellular analysis, where the target cell of interest is limiting and some level of morphometrical or localization evaluation is essential. The Amnis ImageStreamX represents a true crossover platform that can provide high throughput quantitative data assessing both of these attributes, and many more. Described within this application is a wide range of NIH funded research projects that will uniquely benefit from the diverse analytical capabilities of the ImageStreamX. This platform will greatly advance standardization, reproducibility, and throughput, therefore increasing the quality and speed of scientific progress. Additionally, this instrument has the unmatched ability to combine morphological and protein localization differences with flow cytometry fluorescence gating to isolate populations that researchers have never been able to isolate before. Finally, placement of this instrument into a highly respected shared resource lab with dedicated trained staff will ensure its long term operational availability and financial stability.