Laserscanning Mikroskop für Temperatursprung-Experimente

Ebbinghaus小组感兴趣的是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(即与年龄相关的变化)中折叠。该小组研究物理化学因素(如拥挤、限制、水合作用)或生物因素(如伴侣、细胞压力)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，以时空方式直接在细胞环境中解析生物分子的稳定性和动力学。为了建造该仪器，将获得一个共焦显微镜，并配备红外激光，以允许快速和空间分辨率加热样品。该仪器将用于三个研究项目。在第一个项目中，基于蛋白质Barnase特定突变的不同生物传感器被用来量化细胞保持其蛋白质组在空间和时间上折叠的能力。在第二个项目中，所要求的仪器用于测量细胞中SOD1蛋白的致病突变的定位和折叠。进一步，测试了细胞内靶向化学伴侣作为治疗ALS的新药物的效果。第三个研究目标是扩展当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LLP在细胞中在健康和疾病条件下的功能作用。

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.