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

该项目的目的是利用分子生物学多色标记(Multiplex)、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)，对功能核纳米结构进行定量的拓扑分析。建立的方法将进一步发展为病理相关基因区域的例子，慢性粒细胞白血病(CML)相关的bcr区域。此外，通过使用新的“纳米晶体”或“量子点”(QD)来替代目前使用的荧光染料，将改善纳米结构的分析。在另一个应用中，与Cardoso博士(柏林)合作，计划进行荧光共振能量转移(FRET)测量，这将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复制机制中的结构-功能-关系。

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.