Confocal laser-scanning-microscope

共焦激光扫描显微镜

• 批准号: 523980288
• 金额: --
• 依托单位: Ruhr-Universität Bochum
• 依托单位国家: 德国
• 项目类别: Major Research Instrumentation
• 财政年份: 2023
• 资助国家: 德国
• 起止时间: 2022-12-31 至 无数据
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/523980288?language=en
• 关键词: Confocal; laser; scanning; microscope

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).

生物与生物技术学院，特别是所有从事植物模型生物研究的小组，迫切需要一种高分辨率和高灵敏度的共聚焦激光扫描显微镜。对于从事植物研究的小组来说，获得共聚焦显微镜的机会非常有限，以满足蛋白质亚细胞定位、动态结构(如自噬小体、小泡和其他小结构(如加工体、双体、高尔基体等)的活细胞成像)的日益增长的需求。为了在活的植物组织中进行蛋白质-蛋白质相互作用研究(FRET-FLIM)，需要一台最先进的共焦显微镜。所有拟议的项目都旨在(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-蛋白质相互作用的研究。Grefen、Ebert、Krämer、González Fuente和ästün小组的项目也将监测候选蛋白质在不同环境中的动态。此外，Ute Krämer、Suaybústün和Manuel González Fuente的研究小组将在其拟议的项目中进行FRAP实验。为了保证所有拟议项目的成功，拥有最先进的共焦显微镜是至关重要的。该装置的特点是：(I)独特的消除自发荧光的可能性，这对植物研究非常重要；(Ii)通过高分辨率和高灵敏度；(Iii)通过使用多达8条同时独立的激光谱线，允许检测样品中的多个荧光团。有了共振扫描仪，这种显微镜可以高速扫描图像，与旋转的圆盘共焦相当，这对于监测高度移动的结构以及检测容易漂白的弱表达荧光融合蛋白至关重要。集成的薄膜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装置将使我们能够研究活样本中的快速过程(例如，囊泡分选/运输、自噬小体融合事件、信号转导和蛋白质相互作用)。