ShEEP Request for ImageStream®X Mark II Imaging Flow Cytometer

Ralph H Johnson Va Medical Center, Charleston SC

Investigators

Abstract

The goal of this proposal is to install and integrate the ImageStream®X Mark II Imaging Flow Cytometer system in the Cellular and Molecular Evaluation Core (CMEC) Facility within Research Service at the Ralph H. Johnson VA Medical Center. The overarching objective of this core facility is to provide tools and services, including flow cytometry and cell sorting applications, necessary to support and advance research that directly impacts the health of our Veterans. Conventional flow cytometry allows for interrogation of large numbers of cells and quantitative analyses of these populations; however, it does not allow for sub-cellular localization and functional analysis is limited. Conventional microscopy offers morphologic information as well as detailed cellular and subcellular localization data; however, interpretation of microscopic images can be subjective, qualitative, and tedious to quantify. Given the increasing complexity of research questions at both the basic and translational levels, the need for advanced technology that allows for speed, sensitivity, and precision phenotyping and functional analysis is urgent. The ImageStream®X Mark II system represents a first in class, state-of-the-art platform allowing for a powerful combination of multiparametric flow cytometry and imaging, circumventing the limitations of conventional flow cytometry and microscopy to provide a never before available level of speed, sensitivity, quantification, and precision for analysis of animal and patient samples. The ImageStream®X Mark II?s proprietary technology results in superior fluorescence sensitivity for resolution of rare, dim, or small populations and quantitative microscopy for powerful statistical analysis of these populations, allowing for examination of cellular populations, cellular sub-populations, and particles (i.e., exosomes) that simply could not be quantitatively analyzed using standard instrumentation. Our investigators have identified four primary aims that will be accomplished through this groundbreaking technology: 1) Identification of novel cellular sub- populations which contribute to the pathology of disease; 2) Examination of rare events (including stem cells) and small particles (including exosomes) in physiological and pathological conditions; 3) Detection of cellular biomarkers of disease progression and/or clinical response to treatment; and 4) Discovery of novel signaling pathways leading to the development of targeted, personalized, precision therapies. The state-of-the-art technology afforded by the ImageStream®X Mark II Imaging Flow Cytometer System is broadly applicable across the diverse animal- and human-based research efforts of our local VAMC investigators including studies in traumatic brain injury, stroke, spinal cord injury, respiratory illness, cardiovascular disease, immunology, oncology, orthopedic injury, Alzheimer?s disease and mental health disorders. Incorporating the ImageStream®X Mark II Imaging Flow Cytometer System into the established CMEC will greatly strengthen the research capabilities of our station, facilitating basic scientific discovery of mechanisms underlying physiological and pathological processes and translation of these findings to support the rapid implementation of cutting-edge personalized medical treatment to improve quality healthcare for our Nation?s Veterans, a specific directive outlined in the Secretary?s Blueprint for Excellence and Secretary Shulkin?s 10-point plan.