High-Resolution Confocal Laser Scanning Microscope with Spectral Detection

成像是明斯特大学生物科学的核心方法支柱。现代生物传感方法目前在大学中越来越重要。生物传感技术不仅提供细胞、组织和生物体的结构和运动信息，而且增加了对生命系统中局部生理和生化功能的深入了解。最近，通过新任命Markus Schwarzländer为植物生物学和生物技术研究所教授，这将与该研究所的几个研究小组，生物学学院和大学产生协同作用，并支持新的协调举措的发展，这种方法的方向得到了加强。活植物组织将是计划中的基于生物传感的调查的主要焦点。然而，这些技术也将引起几个研究其他系统（如哺乳动物细胞）的研究小组的浓厚兴趣。虽然Schwarzländer实验室在Münster开始之前可以使用合适的仪器，但现在没有共聚焦显微镜系统可以满足访问和技术通用性的要求，以使新研究小组能够运作和未来发展。因此，我们申请了一个倒置的，高分辨率的共焦激光扫描显微镜系统与快速光谱检测。该系统将作为一个灵活的平台，使用荧光蛋白和类似的荧光探针进行活体生物传感。实验将包括样品操作和亚细胞生理学的平行成像活在显微镜舞台上。一个特别的焦点将是在同一个活的样品，这需要灵活的激光激发和快速光谱检测的几个荧光团的定量和并行检测。此外，需要在生命系统中研究亚细胞区室内的结构与功能关系，这将需要用于高分辨率成像的组件。快速和动态的过程需要在对活样品的干扰最小的情况下解决，以获得对亚细胞功能的不受干扰的洞察。研究蛋白质复合物的荧光相关光谱将是对激光器和检测器的额外要求。该系统将在明斯特成像网络内作为分散式仪器运行。这将使其广泛用于高性能荧光显微镜中特别苛刻的应用，并整合大学生物传感研究活动。

Imaging is a central methodological pillar of the biosciences at the University of Münster. Modern biosensing approaches are currently gaining particular importance at the university. Biosensing does not only provide information about structures and movements in cells, tissues and organisms, but adds novel depth of insight about local physiology and biochemical function live in the living system. This methodological direction has recently been strengthened through the new appointment of Markus Schwarzländer as professor at the Institute for Plant Biology and Biotechnology, which will generate synergies with several research groups of the institute, the Faculty of Biology, and the University and support the development of new coordinated initiatives. Living plant tissues will be the main focus of the planned biosensing-based investigations. The techniques will be of strong interest, however, also for several research groups working on other systems, such as in mammalian cells. While the Schwarzländer lab had access to a suitable instrument before starting in Münster, there is now no confocal microscopy system available that fulfills the requirements with respect to access and technical versatility to enable operation and future development of the new research group. Hence, we apply for an inverted, high resolution confocal laser scanning microscopy system with fast spectral detection. The system will serve as a flexible platform for live biosensing using fluorescent proteins and similar fluorescent probes. Experiments will include sample manipulation and parallel imaging of subcellular physiology live on the microscope stage. A particular focus will be the quantitative and parallelized detection of several fluorophores within the same living sample, which requires flexible laser excitation and fast spectral detection. Moreover, investigation of structure-function relationships within subcellular compartments needs to be enabled in living systems, which will require components for high resolution imaging. Fast and dynamic processes need to be resolved with minimal interference with the living sample, to gain unperturbed insight into subcellular functions. Fluorescence correlation spectroscopy to study protein complexes will be an additional requirement for the lasers and detectors. The system will be operated within the Münster Imaging Network as a decentralized instrument. This will enable broad use for particularly demanding applications in high-performance fluorescence microcopy and integrate research activities in biosensing at the university.