Confocal laser scanning microscope

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To investigate neuroscientific questions at the University of Lübeck, we want to apply for a confocal laser scanning microscope. This device will enable imaging experiments that currently cannot be performed on campus. We are a group of researchers working on neurobiological circuits and the interactions of different neural cell types at the interfaces between the periphery and the CNS. Since all applicants work in the same research building, there are optimal conditions for accessibility and use of the instrument. The microscope will replace a 15-year-old confocal microscope at the same location. The proposed microscope will be able to image very fast cellular processes such as the dynamics of the intracellular messengers Ca2+ or cAMP in three dimensions. In addition, different cell markers will be detected in the same sample and quantitative analyses of specific brain structures, e.g. blood vessels, will be carried out with high throughput. The interfaces being studied include cells of the vascular system, specialised glial cells that form part of the blood-brain barrier, and neurons that process peripheral signals such as nociceptive stimuli or hormones. Various research questions dealing with the interaction of peripheral and central signals will be answered with the help of the microscope. For the planned experiments, the microscope must have optimised excitation and emission techniques and reliably distinguish fluorophores with similar fluorescence spectra. Together with a sensitive detection system, this is a big step towards specific signal detection with high sensitivity. Fast imaging is important, firstly to temporally resolve three-dimensional cellular processes in the seconds range, and secondly to enable a high sample throughput with very good spatial resolution. The planned quantitative analyses of many preparations are only possible with high-throughput imaging, and this in turn can only be achieved with high speed. In addition to the sensitivity of the sensors, the scanning speed plays a decisive role in the time required for image acquisition. The microscope we are applying for has a scanning frequency in the kHz range and is thus ideally suited for high-throughput imaging of three-dimensional samples and rapid imaging of dynamic processes in living cells. Overall, we will use the proposed confocal microscope to investigate the subcellular dynamics of CNS cells and to perform high-throughput imaging, ultimately to find new mechanisms and correlations in neurophysiology and neurological diseases.