Laserscanning Mikroskop für Temperatursprung-Experimente

Ebbinghaus小组对研究细胞如何保持其生物分子在空间（即哺乳动物细胞内的空间）和时间（即年龄依赖性变化）中折叠感兴趣。该小组研究物理化学因素（如拥挤，限制，水合）或生物因素（如伴侣，细胞应力）如何调节生物分子折叠和功能。因此，该小组研究了从试管到生物体的不同环境中的生物分子。所要求的仪器结合了高速共聚焦成像和红外激光诱导的温度跳跃，直接在细胞环境中时空解析生物分子的稳定性和动力学。为了构建仪器，将获得共聚焦显微镜并配备红外激光器，以允许快速和空间分辨的样品加热。该仪器将用于三个研究项目。在第一个项目中，基于蛋白质芽孢杆菌RNA酶的特定突变体的不同生物传感器被用于量化细胞在空间和时间上保持其蛋白质组折叠的能力。在第二个项目中，所要求的仪器用于测量细胞中蛋白质SOD 1的致病突变体的定位和折叠。此外，测试了细胞内靶向化学分子伴侣作为针对ALS的新疗法的效果。第三个研究目标是扩展目前关于蛋白质如何在细胞中展开、错误折叠和聚集的观点，以包括液-液相分离过程。这些见解可能会重新定义LLPS在健康和疾病条件下细胞中的功能作用。

The Ebbinghaus group is interested in studying how cells keep their biomolecules folded in space (i.e. spatially inside mammalian cells) and time (i.e. age-dependent changes). The group investigates how physicochemical factors (such as crowding, confinement, hydration) or biological factors (such as chaperones, cell stresses) modulate biomolecular fold and function. Therefore, the group studies biomolecules in different environments ranging from the test tube to the organism. The requested instrument combines high-speed confocal imaging and infrared laser-induced temperature jumps to spatio-temporally resolve biomolecular stability and kinetics directly in cellular environments. To build the instrument, a confocal microscope will be acquired and equipped with an infrared laser to allow for fast and spatially resolved heating of the sample. The instrument will be used in three research projects. In the first project, different biosensors based on specific mutants of the protein barnase are used to quantify the capacity of cells to keep their proteome folded over space and time. In the second project, the requested instrument is used to measure the localization and folding of pathogenic mutants of the protein SOD1 in cells. Further, the effect of intracellular targeted chemical chaperones as new therapeutics against ALS is tested. The third research objective is to extend the current view of how proteins unfold, misfold and aggregate in cells to include liquid-liquid phase separation processes. The insights will potentially redefine the functional role of LLPS in cells in health and disease conditions.