Atomic Force Microscope with Fluorescence Microscope for simultaneous measurements

带有荧光显微镜的原子力显微镜可同时进行测量

• 批准号: 441175622
• 金额: --
• 依托单位: Universität zu Lübeck
• 依托单位国家: 德国
• 项目类别: Major Research Instrumentation
• 财政年份: 2020
• 资助国家: 德国
• 起止时间: 2019-12-31 至 无数据
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/441175622?language=en
• 关键词: Atomic; Force; Microscope; Fluorescence; simultaneous

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.

内皮细胞形成血管的最内层，并暴露于流动血液的剪切力。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机械性能和灵活的反应，以不同程度的剪切力。特别地，内皮细胞的外壳可以在功能上分为糖萼和皮质，它们都是高度柔性和反应性的。软和硬内皮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扩张剂一氧化氮（NO）的释放和生物利用度：软内皮细胞的表面因流动的血液而更变形，并比硬内皮细胞分泌更多的NO。因此，“刚度”是反映细胞生理状态的机械特性，并且可以被视为细胞功能的标志。我们可以发现，长期僵硬的内皮导致内皮功能障碍，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，我们的小组感兴趣的是分析和量化的内皮细胞的力学特性，他们的功能，功能障碍和潜在的细胞机制。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机械特性。通过这种技术，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） 是单细胞功能的标志物7） 与血管活性物质的生物利用度相关8） 我们未来几年的目标是阐明细胞力学的潜在分子机制，并将这些知识转化为转化方法。因此，将进一步表征内皮表面的机械性质。特别是，内皮糖萼是我们项目的重点。以下几点具有很高的优先级：4） 细胞力学与血管炎症5） 在疾病模型（肾脏疾病中的尿毒症、急性心肌梗死、糖尿病）背景下内皮硬度的病理生理学变化6） 基因和高血压为了成功地实施这些项目，我们将量化机械性能并使用AFM对内皮细胞进行成像。此外，将采用单细胞力光谱法来量化细胞（例如单核细胞-内皮细胞）之间的粘附力。在我从明斯特大学第二生理学研究所转到吕贝克大学生理学研究所以及一个新的研究小组的相关发展之后，收购拟议的Nanowe 4对于项目的成功延续至关重要。