PROJECT SUMMARY/ABSTRACT The University of Iowa Flow Cytometry Facility was established in 1979 and currently serves 128 principal investigators spanning 31 departments across four colleges. This group continuously receives NIH support for their work on diverse biomedical research projects that advance our understanding of biological processes that are essential for human health and identify novel methods to detect and treat a wide variety of diseases. Recent advances in flow cytometry, microscopy, and single-cell sequencing have increasingly enabled researchers to identify specific cell populations. Consequently, our investigators require cutting-edge instrumentation that can accommodate a wide range of experimental conditions to address new research questions that arise due to these expanding technologies. Currently, our Facility operates six heavily used benchtop flow cytometers. However, the Facility lacks an imaging cytometer, which harnesses the high throughput, statistically robust data acquisition of a flow cytometer with the imaging and morphological capabilities of microscopy. In the absence of this, investigators must run samples on a flow cytometer to generate quantitative data and use the rest of their sample, or a duplicate of their sample, to generate qualitative images on a microscope. This workflow is incompatible with rare cell populations, limits the rigor and reproducibility of experiments due to different labeling schemes, and is costly and time consuming. The nearest publicly accessible imaging cytometer is over 200 miles away. This lack of accessibility to an imaging cytometer has a significant adverse impact on the ability of our investigators to obtain detailed images and corresponding robust statistical data to assess a wide range of biological questions. Thus, there is an urgent need to acquire an imaging cytometer in the Flow Cytometry Facility at the University of Iowa so investigators can expand their capacity to understand cell morphology, identify subcellular localization, assess protein- protein interactions, and distinguish specific cell staining from background in rare cell populations in a quantitative manner, which will enable them to advance their research in new directions. This proposal requests funds for a Cytek Amnis ImageStreamX MkII imaging cytometer to provide NIH-funded investigators and other researchers at the University of Iowa with access to an instrument that can generate both qualitative and quantitative data, thus reducing the time needed for data acquisition, sample degradation, and sample-to-sample variation in the case where different samples are run using flow cytometry vs microscopy. This will allow for continued support and enhancement of research projects carried out at the University of Iowa and in the state of Iowa, many of which are part of clinical trials or crucial pre-clinical or basic science studies, and will ensure the Facility continues to meet its overall goal of providing sta...