Three-dimensional imaging of cells and tissues with ultrahigh spatiotemporal resolution by lattice light sheet microscopy

将生物分子的高度动态组织分解为功能单元是从机理上理解生物过程的关键。凭借其与生命细胞成像的兼容性，其非常高的灵敏度和选择性，以及其多功能的工具和报告探针分子和细胞功能，荧光显微镜提供了独特的可能性，以应对这一挑战。通过不同的所谓的超分辨率成像技术克服衍射极限分辨率，刺激了细胞分子过程的荧光显微镜的巨大发展。在奥斯纳布吕克大学生物系，这些令人兴奋的发展很早就被用来解决非常不同的生物背景下的基本细胞生物学问题。在生物物理学部专业知识的支持下，我们专注于基于单分子定位的成像技术，我们认为这是最适合解开活细胞中分子过程的时空动力学。这些努力是由一个联合研究倡议，合作研究中心944“生理学和细胞微区室的动力学”，在2011年起飞的促进和协调。目前，来自Osnabrück的九个小组和来自Münster的四个小组从不同的生物学角度合作研究了这个CRC，研究了细胞中的生物分子如何动态地自组织成功能性微区室。该CRC的重要资产是我们在其第一个资助期内建立的专用先进光学显微镜设施。方法开发和应用的高度多样化的生物学问题和模型系统的密切接口产生了活细胞单分子定位显微镜的独特的专业知识。

Resolving the highly dynamic organization of biomolecules into functional units is the key to mechanistic understanding of biological processes. With its compatibility with life cell imaging, its very high sensitivity and selectivity as well as its versatile tools and reporters to probes mo-lecular and cellular functions, fluorescence microscopy offers unique possibilities to tackle this challenge. Overcoming the diffraction limited resolution by different so-called superresolution imaging techniques has stimulated tremendous developments in fluorescence microscopy of molecular processes in cells. At the Biology Department of the University of Osnabrück, these exciting developments were early on taken up to tackle fundamental cell biological questions in very different biological context. Supported by the expertise of the Division of Biophysics, we fo-cused on single molecule localization-based imaging techniques that we felt were most suitable to unravel the spatiotemporal dynamics of molecular processes in living cells. These efforts are fostered and coordinated by a joint research initiative, the Collaborative Research Center 944 “Physiology and Dynamics of Cellular Microcompartments” that took off in 2011. Currently, nine groups from Osnabrück and four groups from Münster collaborate in this CRC from different bio-logical perspectives on the overarching question how biomolecules in cells dynamically self-organize into functional microcompartments. Important asset of this CRC is a dedicated advanced light microscopy facility that we established during its first funding period. Close interfacing of method development and application to highly diverse biological problems and model systems has yielded a unique expertise in live cell single molecule localization microscopy.