Atomic Force Microscope with Fluorescence Microscope for simultaneous measurements

内皮细胞形成血管的最内层，并暴露在血流的剪切力下。这种“战略位置”使血管内皮细胞能够控制血管的功能。因此，内皮细胞的表面可以改变其力学性能，并对不同程度的剪切力做出灵活的反应。特别是内皮细胞的外壳可以分为糖萼和皮质，它们都具有高度的柔韧性和反应性。软质和硬质内皮细胞表面之间的变化具有高度的生理相关性，并调节血管活性物质（如血管舒张剂一氧化氮（NO））的释放和生物利用度：软质内皮细胞表面受血流的影响更变形，比硬质内皮细胞分泌更多的NO。因此，“刚度”是一种反映细胞生理状态的机械特性，可以看作是细胞功能的标志。我们可以证明，长期僵硬的内皮会导致内皮功能障碍，并有助于心血管疾病的发展，如动脉粥样硬化和高血压。多年来，我们的团队对内皮细胞的力学特性，功能，功能障碍和潜在的细胞机制的分析和量化感兴趣。通过使用原子力显微镜（AFM），可以扫描内皮表面以量化细胞的力学特性。通过这项技术，我们可以证明机械刚度(5)取决于皮质细胞骨架和膜蛋白（6)是单个细胞功能的标志（7)与血管活性物质的生物利用度相关）(8)可以在治疗上进行操作）。您未来几年的目标是阐明细胞力学的潜在分子机制，并将这些知识应用于翻译方法。因此，内皮表面的力学性能将进一步表征。特别是内皮糖萼是我们项目的重点。4)细胞力学和血管炎症5)疾病模型（肾病尿毒症、急性心肌梗死、糖尿病）背景下内皮刚度的病理生理变化6)基因和高血压为了成功开展这些项目，我们将量化内皮细胞的力学特性并使用AFM成像。此外，单细胞力光谱将用于量化细胞之间的粘附力（例如单核细胞-内皮细胞）。在我从明尼苏达大学第二生理学研究所转到伊利诺斯大学<s:1>贝克大学第二生理学研究所并成立了一个新的研究小组之后，收购拟议的Nanowizard 4对于项目的成功延续是必不可少的。

Endothelial cells form the innermost layer of blood vessels and are exposed to the shear forces of the streaming blood. This ‘strategic positions’ enables the vascular endothelial cells zu control the function of the vessel. Thereby, the surface of endothelial cells can change their mechanical properties and flexible react to different graduations of shear forces. Particularly, the outer shell of endothelial cells can be functionally divided in glycocalyx and cortex which are all highly flexible and reactive. The change between a soft and stiff endothelial surface is of high physiological relevance and regulates the release and bioavailability of vasoactive substances such as the vasodilator nitric oxide (NO): the surface of soft endothelial cells is more deformed by the streaming blood and secrete more NO than stiff endothelial cells. Thus, ‘stiffness’ is a mechanical property which reflects the physiological state of cells and can be seen as hallmark for the function of cells. We could show that a chronically stiff endothelium leads to endothelial dysfunction and contributes to the development of cardiovascular pathologies such as atherosclerosis and hypertension.For many years our group is interested in the analysis and quantification of the mechanical properties of endothelial cells, their function, dysfunction and the underlying cellular mechanisms. By using an Atomic Force Microscope (AFM) it is possible to scan the endothelial surface to quantify the mechanical properties of the cells. With this technique we could show that mechanical stiffness 5) Depends on the cortical cytoskeleton and membrane proteins6) Is a marker for the function of single cells7) Correlates with the bioavailability of vasoactive substances8) Can be manipulated therapeuticallyOur goal for the next years is to elucidate the underlying molecular mechanisms of cell mechanics and to implement this knowledge into a translational approach. Therefore, the mechanical properties of the endothelial surface will be further characterized. In particular, the endothelial glycocalyx is in the focus of our projects. The following points have a high priority:4) Cell mechanic and vascular inflammation5) Pathophysiological changes of endothelial stiffness in the context of disease models (uremia in kidney diseases, acute myocardial infarction, diabetes)6) Genes and hypertensionTo carry out these projects with success, we will quantify the mechanical properties and image endothelial cells with the AFM. In addition, single-cell-force spectroscopy will be employed to quantify adhesion forces between cells (e.g. monocyte – endothelial cell). After my change from University of Münster, Institute of Physiology II to University of Lübeck, Institute of Physiology and the associated development of a new research group, the acquisition of the proposed Nanowizard 4 is imperative for the successful continuation of the projects.