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Special methodical expertise

Integrins and their ligands are prominent players regulating interactions of the extracellular microenvironment and cellular functions. Sensing the biomechanic changes of microenvironment integrins bidirectionally transmit conformational changes across the membrane. On demand integrin and their ligand modulate the structure and function of their microenvironment. The understanding of integrin ligand-dependent mechanotranduction elucidates e.g. tumorgensis, stem cell differentiation, and hemostasis. Especially the analysis of prothrombotic mutations that are risk determinants of acute coronary syndromes needs special flow models to simulate biomechanic stress by blood flow and real time fluorescent image acquisitions systems to monitor molecular and structural changes:

• By flow simulation in an in vitro flow model or rheometer we elucidate the conformational and functional changes of the integrins and ligands. In a blood flow model we use human platelets or cell lines expressing integrins to measure fibrillogenesis of fibronectin in response to mechanotransduction and biomechanic stress.
• Multiparametric confocal laser scanning microscopy and three dimensional imaging (Figure 1) is used to measure fibrillogensis of fibronectin (protein unfolding) by e.g. disappearance of fluorescence by fluorescence resonance energy transfer (FRET).
• Multiparametric flow cytometrie and fluorescence imaging are used to quantify the integrin expression, their function and activation as well as their ligand binding on a cellular level.

Figure Legend:
A: Three dimensional thrombogenesis upon in vitro blood flow acquired by confocal laser scanning microscopy. Integrin and ligand are responsible for thrombus-formation. Endpoint after 10 minutes of blood flow is shown as an overview (left) and a magnification (right). A movie of thrombogensis could be seen on:
 http://bv.acs.uni-duesseldorf.de/projects/thrombo.html

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