Lightsheet-Microscopy-System

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A white light laser Lightsheet microscope is proposed for the interdisciplinary research platform "Multiparametric Bioimaging and Cytometry" (MPBIC) of the Medical Faculty of the Otto-von-Guericke-Universität Magdeburg. The MPBIC platform is the central research infrastructure for light microscopy and flow cytometry approaches in the medical faculty within the research focus "Immunology and molecular medicine of inflammation". The imaging techniques currently available in the MPBIC (multiphoton-microscopy, confocal microscopy, wide-field microscopy) allow for the visualization of small samples or small sections of larger samples with high resolution. Larger samples have to be cut into thin sections due to the restricted penetration and emission depth of the exciting or the emitted light, which is leading to preparation artefacts and sample loss. However, various research objectives require the analyses of cell infiltrates or tissue maladaptation/structural alterations in the intact three-dimensional tissue context, which can be not addressed with thin sections (or only incompletely). The multi-photon intravital microscopy, which is a well-established procedure in the MPBIC platform, allows for the analysis of cell dynamics in the three-dimensional tissue but is limited to small sample regions despite the high penetration depth of the exciting laser. Consequently, this method is only limitedly useful for research projects addressing inflammatory processes, which are not homogeneously dispersed, but occurring in form of infiltration or infection spots. Recently, various methods of whole-mount staining and subsequent tissue clearing have been developed that enable the detection of structures in whole intact organs with fluorochrom-conjugated antibodies. Since, in practice, the combination of clearing with the scanning techniques available in the MPBIC leads to extremely long measuring times, this procedure is unsuitable for a broad application in the faculty. Therefore, the visualization and analysis of whole intact organs or tissues is not possible with the currently present microscopes or in a very restricted number. The herein proposed lightsheet-microscope will cover the increasing need in the faculty for fast 3D-analyses of large and intact biological samples (organs of experimental animals, organoids, and human biopsies) from the macro into the micro level. This will enable the detection of anatomical-structurally alterations/modifications in the tissue level up to infiltrations of immune cells, or structural alterations in inhomogeneously dispersed inflammation- or infection spots with cellular-subcellular resolution, which is indispensable for a number of promising running and planned research projects.