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论支柱。现代生物传感方法目前在该大学获得了特别重要的地位。生物传感不仅提供关于细胞、组织和生物体的结构和运动的信息，而且增加了对生活在生命系统中的局部生理和生化功能的新的深入了解。最近，通过任命Markus Schwarzländer为植物生物学和生物技术研究所的教授，这一方法方向得到了加强，这将与该研究所、生物系和该大学的几个研究小组产生协同效应，并支持制定新的协调一致的举措。活的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，研究其他系统的几个研究小组也会对这些技术产生浓厚的兴趣，比如在哺乳动物细胞中。虽然施瓦茨兰德实验室在明斯特开始之前可以使用合适的仪器，但现在没有可用的共焦显微镜系统来满足访问和技术通用性方面的要求，以支持新研究小组的运作和未来的发展。因此，我们申请一种具有快速光谱检测功能的倒置、高分辨率共聚焦激光扫描显微镜系统。该系统将成为使用荧光蛋白和类似荧光探针进行活生物传感的灵活平台。实验将包括样品处理和在显微镜舞台上现场直播的亚细胞生理学的并行成像。一个特别的重点将是对同一活样本中的几个荧光团进行定量和并行检测，这需要灵活的激光激发和快速的光谱检测。此外，需要在活体系统中研究亚细胞间隔内的结构-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本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。研究蛋白质复合体的荧光相关光谱将是对激光器和检测器的额外要求。该系统将作为一个分散的仪器在明斯特成像网络内运行。这将使高性能荧光显微镜中特别苛刻的应用得到广泛应用，并整合该大学在生物传感方面的研究活动。

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.