Optical tweezers coupled to a confocal fluorescence microscope and microfluidic Flow Cell

维尔茨堡大学的鲁道夫·维尔绍中心拥有最先进的生物样本成像基础设施。我们可以分辨从Å分辨率的样品，无缝地移动到纳米范围，最终上升到整个生物体。为了实现这种高动态分辨率范围，我们采用了x射线晶体学，低温电镜，超分辨率荧光显微镜，原子力显微镜和共聚焦显微镜等技术，仅举几例。所提出的超高分辨率光镊与高性能共聚焦荧光显微镜和四向微流体层流细胞仪相结合，是一种将光镊力显微镜与共聚焦荧光显微镜相结合的独特设备，是实时单分子分析和操作的理想选择。它将为单分子分辨率下实时研究蛋白质/核酸复合物的功能和机制提供一种重要的新技术，并将我们的成像能力提升到一个新的水平。在本提案中，我们设想解决和解决维尔茨堡大学的几个重要研究项目，包括RNA代谢，G4四联体和核苷酸切除DNA修复过程。该仪器提供的数据将大大提高我们对这些不同研究领域分子过程的理解，否则这些研究领域将无法实现。

The Rudolf Virchow Center at the University of Würzburg harbors state of the art infrastructure for imaging of biological samples. We can resolve samples ranging from Å resolution, seamlessly moving into the nm range and finally going up into the entire organism. To achieve this highly dynamic resolution range we harbor techniques like x-ray crystallography, cryo-EM, super resolution fluorescence microscopy, atomic force microscopy and confocal microscopy just to name a few. The proposed ultra-high-resolution optical tweezers coupled to a high-performance confocal fluorescence microscope and a 4-way microfluidics laminar flow cell instrument is a unique device that is capable of combining force microscopy using optical tweezers with confocal fluorescence microscopy which is ideal for single molecule analysis and manipulation in real time. It will provide an important new and powerful technique to study the function and mechanisms of proteins/nucleic acid complexes at single molecule resolution in real time and bring our imaging capabilities to the next level. In this proposal we envision to tackle and address several important research projects at the University of Wuerzburg that comprise the processes of RNA metabolism, G4 quadruplexes and nucleotide excision DNA repair. The data delivered by the proposed instrument will significantly enhance our understanding of molecular processes in these different research areas which could otherwise not be realized.