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

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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.