• Categories:FCS Technology for Molecular Interaction Analysis
• Time of issue:2023-05-24 14:24
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(Summary description)Jan Ellenberg's group at the European Molecular Biology Laboratory has developed a high-throughput fluorescence correlation spectroscopy (HT-FCS) technique to study the dynamics of protein molecular networks in living cells. motility, interactions, etc. The manual data collection and analysis methods of traditional FCS experiments are only suitable for analyzing the properties of a few target molecules in a few cells in a relatively short period of time. The HT-FCS technology developed by the authors of this thesis enables automatic screening of target cells and automatic acquisition and analysis of fluorescence autocorrelation (FACS) and cross-correlation (FCCS) spectral data at characteristic spatial loci and time points of individual living cells. Applying HT-FCS technology, the research team has performed 60,000 FCS experiments in 10,000 human-derived cells and analyzed the dynamic molecular mechanisms of 53 cellular nuclear proteins. Based on the multiple experimental parameters (concentration, motility, interactions, etc.) obtained from the FCS experiments, these nuclear proteins can be classified into three categories based on their binding to chromatin: 1) nuclear proteins bound to histones to form chromatin complexes; 2) nuclear proteins not bound to chromatin; and nuclear proteins forming molecular complexes of uneven size but not bound to chromatin. In addition, the dynamic changes of Aurora B Kinase and INCENP protein molecules during mitosis were investigated by HT-FCS technique, and Aurora B Kinase and INCENP are crucial for the regulation of chromatin mobile complex (CPC) formation. HT-FCS technology is important for the study of dynamic molecular mechanisms in living cells and drug discovery.