Confocal laser-scanning-microscope

生物和生物技术学院，特别是所有研究植物模式生物的小组，迫切需要一台具有高分辨率和灵敏度的共聚焦激光扫描显微镜。对于研究植物的小组来说，使用共聚焦显微镜是非常有限的，并且为了满足对蛋白质的亚细胞定位、动态结构如自噬体、囊泡和其他小结构（例如，加工体、两性体、高尔基体等）和蛋白质-蛋白质相互作用的研究（FRET FLIM）在活的植物组织，一个国家的最先进的共聚焦显微镜是必需的。所有拟议的项目旨在（i）在不同条件下定位或共定位荧光标记的蛋白质，以及（ii）使用FRET-FLIM和/或BiFC进行蛋白质-蛋白质相互作用研究。Grefen、Ebert、Krämer、González Fuente和Üstün小组的项目也将监测候选蛋白在不同环境中的动态。此外，Ute Kramer、Suayb Üstün和Manuel González Fuente的研究小组将在其拟议的项目中进行FRAP实验。为了保证所有项目的成功，获得最先进的共聚焦显微镜至关重要。所提出的装置的特征在于（i）消除自发荧光的独特可能性，这对于植物研究非常重要，（ii）通过高分辨率和灵敏度，以及（iii）通过使用多达8个同时独立的激光线，其允许检测样品中的多个荧光团。与共振扫描仪，这种显微镜可以扫描图像在高速，可与旋转盘共聚焦，这是必不可少的监测高度移动的结构，以及检测弱表达的荧光融合蛋白，易于漂白。集成FLIM单元是该设备的独特之处，简化了测量前和测量后的数据分析。使用FLIM单元将使我们能够研究活体样品中的快速过程（例如，囊泡分选/转运、自噬体融合事件、信号转导和蛋白质-蛋白质相互作用）。

The Faculty of Biology and Biotechnology, specifically all groups working on plant model organisms, urgently need a confocal laser scanning microscope with high resolution and sensitivity. Access to confocal microscopy is very limited for the groups working on plants and to meet the increased demand for subcellular localization of proteins, live cell imaging of dynamic structures such as autophagosomes, vesicles and other small structures (e.g., processing bodies, amphisomes, Golgi etc.) and protein-protein interaction studies (FRET-FLIM) in living plant tissues, a state-of-the art confocal microscope is required. All proposed project aim (i) to localize or co-localize fluorescently tagged proteins under different conditions, and (ii) to perform protein-protein interaction studies using FRET-FLIM and/or BiFC. Projects from the Grefen, Ebert, Krämer, González Fuente and Üstün groups will also monitor dynamics of candidate proteins in different contexts. Moreover, the research groups of Ute Krämer, Suayb Üstün and Manuel González Fuente will perform FRAP experiments in their proposed projects. To guarantee the success of all proposed projects, it is vital to have access to a state-of-the art confocal microscope. The proposed device is characterized (i) by unique possibilities for the elimination of autofluorescence, which are of great importance for plant research, (ii) through high resolution and sensitivity and (iii) by using up to 8 simultaneous independent laser lines which allows the detection of multiple fluorophores in a sample. With the resonant scanner this microscope can scan images at high speed, comparable to that of a spinning disc confocal, which is essential to monitor highly mobile structures as well as detect weakly expressed fluorescent fusion proteins that are prone to bleaching. The integrated FLIM unit is unique for this device, simplifying the pre- and post-measurement data analysis. Using the FLIM unit will allow us to investigate fast processes in living samples (e.g., vesicle sorting/transport, autophagosome fusion events, signal transduction and protein-protein interactions).