Atomic Force Microscope with Fluorescence Microscope for simultaneous measurements

内皮细胞形成血管的最内层，并暴露在流动的血液的剪切力下。这种“战略位置”使血管内皮细胞能够控制血管的功能。因此，内皮细胞表面可以改变其力学性质，并对不同程度的剪切力做出灵活的反应。特别是，内皮细胞的外壳在功能上可以分为糖萼和皮质，它们都具有高度的弹性和反应性。柔软和僵硬的内皮细胞表面之间的变化具有很高的生理相关性，并调节血管活性物质的释放和生物利用度，如血管扩张剂一氧化氮(NO)：柔软的内皮细胞表面被流动的血液变形得更多，分泌的NO比僵硬的内皮细胞更多。因此，“僵硬”是反映细胞生理状态的一种力学性质，可视为细胞功能的标志。我们可以证明，长期僵硬的内皮细胞会导致内皮功能障碍，并有助于动脉粥样硬化和高血压等心血管疾病的发展。多年来，我们的团队对分析和量化内皮细胞的机械特性、功能、功能障碍和潜在的细胞机制感兴趣。通过使用原子力显微镜(AFM)，可以扫描内皮细胞表面来量化细胞的机械性能。利用这项技术，我们可以证明机械硬度取决于皮质细胞骨架和膜蛋白6)是单细胞功能的标志7)与血管活性物质的生物利用度相关8)可以进行治疗我们未来几年的目标是阐明细胞力学的潜在分子机制，并将这一知识转化为翻译方法。因此，将进一步表征内皮细胞表面的力学性能。特别是，内皮细胞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(肾脏疾病中的尿毒症、急性心肌梗死、糖尿病)背景下内皮细胞僵硬的病理生理变化6)基因和高血压为了成功地实施这些项目，我们将用原子力显微镜量化内皮细胞的机械特性和图像。此外，还将使用单细胞力谱来量化细胞(例如单核细胞-内皮细胞)之间的粘附力。在我从明斯特大学生理学研究所II改为L大学生理学研究所，并建立了一个新的研究小组后，收购拟议中的Nanowizard 4对于项目的成功继续是必不可少的。

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.