4-Laser Cell Sorter

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With my appointment at the chair of Pharmaceutical Biology at the Friedrich-Alexander-University Erlangen-Nürnberg, I would like to initiate a scientific reorientation of the research focus. In the coming years, we will be working intensively on bacterial infectious diseases, which represent one of the top three threats to humanity. We are investigating how bacteria communicate with each other and modulate host cells by means of cell-derived vesicles. These extracellular vesicles are fundamental cell communicators and relevant in various pathophysiologies. Our fundamental findings contribute to the development of urgently needed new therapeutic options in the field of bacterial infections and inflammatory diseases. The laboratories at the chair will be extensively renovated in 2022/23 to create an optimal research infrastructure. Following these refurbishments, I plan to establish an independent S2 area in which infection models on pathogens will be optimised and evaluated. For this purpose, I would like to equip a measuring area with a confocal fluorescence microscope and a flow cytometer that is also accessible to other users. I am submitting separate but parallel applications for these two major research instruments and I am asking for a joint review, as the devices as these devices are a common integral part of the S2 infrastructure to be created at the chair. The confocal microscope is primarily used for the spatial-temporal visualisation of infection models, while flow cytometry is used to quantify the cellular samples. An areal separation of these two devices would entail uncertainties and instabilities during the transport of the cell samples and would therefore mean a significant reduction in scientific results and impair the development of new anti-infective therapies. The planned benchtop flow cytometer has a wide applicability and is equipped with four lasers and thus the possibility to measure at least 15 fluorescent colours in parallel via two scattered light detectors (side and forward scattering). The lasers cover the wavelengths 405 nm (violet), 488 nm (green), 561 nm (yellow-green) and 638 nm (red) to measure most typical fluorophores and compensations. The possibility to directly measure vesicles in the size range around 200 nm via a violet laser allows to distinguish them from cells and cell debris. All fluorescence must be measurable with a resolution of 10-30 MESF (Molecules of Equivalent Soluble Fluorochromes) to achieve the highest sensitivity. The device shall provide stable and bubble-free sorting of live bacteria in well plates. In addition to the FACS device, a powerful data acquisition PC is also required, as well as a software dongle that can be used to evaluate the analyses on individual workstation computers. The device is installed in a separate, dark room.