Analysis of the 3D-nuclear-nanostructure in human Cells using Fluorescence Detection of Multiplex Dyes

使用多重染料荧光检测分析人体细胞中的 3D 核纳米结构

• 批准号: 5107134
• 负责人: Professor Dr. Christoph Cremer
• 金额: --
• 依托单位: Institut für Molekulare Biologie gGmbH (IMB)
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 1998
• 资助国家: 德国
• 起止时间: 1997-12-31 至 2006-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5107134?language=en
• 关键词: Analysis; 3D; nuclear; nanostructure; human

The aim of the proposed project is the quantitative topological analysis of functional nuclear nanostructures, using molecular biological multi colour labeling (Multiplex), high resolution microscopy and digital image analysis (distance resolution goal between four labeled DNA-sequences = 50 nm). The established methods will further be developed for the example of a pathologically relevant gene region, the chronic myelogenous leukemia (CML) correlated BCR-region. Furthermore, nano structure analysis will be improved by the use of new "Nano-Crystals" or "Quantum Dots" (QD's) as a substitute for the presently used fluorochromes. In another application, in cooperation with the group of Dr. Cardoso (Berlin) fluorescence resonance energy transfer (FRET) measurements are planned which should give an insight into the structure function relation of DNA replication with a 3D-resolution of up to 2-8 nm on the single molecule level. The aim is the study of the structure-function-relation in the DNA replication machinery of living cells.

该项目的目标是利用分子生物多色标记（多重）、高分辨率显微镜和数字图像分析（四个标记 DNA 序列之间的距离分辨率目标 = 50 nm）对功能性核纳米结构进行定量拓扑分析。已建立的方法将进一步针对病理相关基因区域（慢性粒细胞白血病 (CML) 相关 BCR 区域）的示例进行开发。此外，通过使用新的“纳米晶体”或“量子点”（QD）作为目前使用的荧光染料的替代品，纳米结构分析将得到改进。在另一项应用中，我们与 Cardoso 博士（柏林）团队合作计划进行荧光共振能量转移 (FRET) 测量，该测量可在单分子水平上以高达 2-8 nm 的 3D 分辨率深入了解 DNA 复制的结构功能关系。目的是研究活细胞 DNA 复制机制的结构-功能关系。