5-Laser Flow Cytometer

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Flow cytometry constitutes a versatile technology for the analysis of single cells. It is based on labeling of cells with a combination of fluorophores followed by hydrodynamic separation and analysis of laser-induced fluorescence. The power of flow cytometry derives from a large variety of fluorescent labeling techniques, ranging from fluorochrome-conjugated antibodies directed against cellular surface as well as intracellular structures, including proteins and nucleic acids, via uptake of fluorescent metabolite analogs to genetically encoded reporters for defined cellular states, such as activation of signaling pathways, cell cycle, cell death or senescence. Furthermore, flow cytometry is ideally suited to analyze complex cell populations in a short amount of time. Accordingly, flow cytometry has emerged as a standard method in cellular and molecular Immunology as well as in adjacent fields like Cell Biology and Physiology. Progress in flow cytometry critically needs to keep track with the increasingly detailed description of cell types and their characteristics. In this proposal, an analytical flow cytometer is requested, capable of rapid and sensitive parallel analysis of up to 21 parameters (including 19 fluorescent parameters) in a 5-laser setup. This multiparameter 5-laser flow cytometer will become part of the flow cytometry core facility of the Faculty of Medicine as well as part of a new research focus of Molecular Immunology of the Faculty of Biology and Chemistry to foster the university’s area of potential “Infections, Inflammation and Active Substances” and its accent area “RNA Biochemistry”. It will primarily serve to advance projects aimed at characterizing development and function of immune cells. Specifically, one project is dedicated to quantitatively map cell cycle dynamics of developing T cells in the thymus, employing combinations of cell surface markers, analysis of DNA replication speed and transgenically encoded fluorescent cell cycle indicators. Additional projects focus on the dynamic metabolic changes in Mucosa-associated invariant T cells and their function in physiology and pathophysiology of the lung as well as lung remodeling upon insult in general. These projects are based on the analysis of complex subpopulations employing metabolic markers as well as the activation of critical signaling pathways. Further projects focus on the analysis of microRNA activity in the context of T cell receptor-dependent signaling pathways, characterization of immune cell subpopulations in the epidydimis as well as in multiple-sclerosis-related pathology. Administrative integration of the requested multiparameter 5-laser flow cytometer into a core facility structure will ensure its efficient and interdisciplinary use.