Atomic Force Microscope

对与生物细胞和组织功能相关的力学性质的详细研究极大地加深了我们对与健康细胞功能相关的复杂调控的理解，也使我们对几种病理情况下的复杂调控有了更深入的了解。为了全面了解细胞和组织力学，我们的团队专注于对细胞内以及细胞-细胞和细胞-基质界面上的粘弹性性质和力的产生进行详细的量化。由于我最近从明斯特搬到了哥廷根，我需要购买一台AFM，以便在新的主办机构建立这样的项目。由于现代生物物理问题需要对所研究的生物系统有详细的了解，我们需要同时获得细胞力学测量，同时获得关于通过荧光显微镜研究的系统的结构和动力学信息。由于我们对具有相关3D扩展的生物系统感兴趣，我们将把AFM连接到研究所现有的旋转圆盘显微镜上。虽然旋转圆盘的振动可能会降低原子力显微镜的空间分辨率，但不影响我们在组织力学中的应用。3D旋转圆盘显微镜和AFM力学的独特组合，以及基于AFM的粘附性测量，将使我们能够直接描述单个细胞中细胞内和皮质机械性能之间的相互作用，以及重组组织和有机体内的相互作用。除了间期细胞，我们还将研究有丝分裂细胞和正在进行减数分裂的卵母细胞。在进一步的方向，我们将研究组织的粘弹性性质，如癌球体，或小器官，如腔形成上皮腺泡。从AFM测量获得的机械信息将与从光片显微镜获得的组织流动测量相结合，以通过流体动力学方法直接描述组织动力学。在这里，不仅粘弹性属性是相关的，我们还需要确定细胞整合到组织中所使用的细胞-细胞粘附力。在另一个方向，我们研究肌肉组织的机械稳态，我们发现来自健康受试者和患有dystrophin蛋白突变的患者的重组肌肉组织的机械稳态存在显著差异，这会导致严重的肌肉营养不良。除了这些主要方向外，新的显微镜还将用于局部合作，研究细胞-细胞连接中的机械效应(Andreas Janshoff)和中间细丝对皮质硬度的影响(Sarah Köster)。在进一步的方向上，我们的目标是由DNA-折纸(Jörg Enderlein)形成的成像结构。在这个项目中，旋转盘部件将被关闭以避免额外的振动。

The detailed investigation of the mechanical properties relevant for biological cell and tissues function has greatly enhanced our understanding of the complex regulation relevant in both, healthy cell function but also in several pathological situations. To gain a full picture of cell and tissue mechanics, our team has its focus on a detailed quantification of the viscoelastic properties and the force generation within cells and at the cell-cell and cell-substrate interface. Due to my recent move from Münster to Göttingen, I need to purchase an AFM to establish such projects in the new host institute. As modern biophysical questions require detailed knowledge about the biological systems studied, we need to simultaneously acquire the cell mechanical measurements while gaining structural and dynamical information about the system investigated via fluorescent microscopy. Since we are interested in biological systems with a relevant 3D extension, we will attach the AFM to a spinning disk microscope that is present in the institute. Although the vibration of the spinning disk might reduce the spatial resolution of the AFM, our application in tissue mechanics is not affected. The unique combination of 3D spinning disk microscopy and AFM mechanics, as well as AFM based adhesion measurements will allow us to directly describe the interactions between intracellular and cortical mechanical properties in single cells, but also in reconstituted tissue and organoids. Beside interphase cells, we will also study mitotic cells, and oocytes undergoing miosis. In a further direction we will investigate the viscoelastic properties of tissue, such as cancer spheroids, or small organoids such as lumen forming epithelial acini. The mechanical information obtained from the AFM measurements will be combined with tissue flow measurements obtained from light sheet microscopy, to directly describe tissue dynamics by a hydrodynamical approach. Here not only the viscoelastic properties are relevant, we also need to determine the cell-cell adhesion forces used by the cells to integrate into the tissue. In another direction we investigate the mechanical homeostasis of muscle tissue, where we see significant differences in the mechanical homeostasis of reconstituted muscle tissue from healthy subjects and patients suffering for a mutation in the dystrophin protein, leading to severe muscular dystrophies. Besides these main directions, the new microscope will be used in local cooperation to study the mechanical effect in cell-cell junctions (Andreas Janshoff) and the influence of intermediate filaments on the cortical stiffness (Sarah Köster). In a further direction we aim at imaging structures formed by DNA-origami (Jörg Enderlein). For this project the Spinning-Disk component will be turned off to avoid additional vibrations.