3D-STED Microscope

3D STED 显微镜

• 批准号: 449737235
• 金额: --
• 依托单位: Johannes Gutenberg-Universität Mainz
• 依托单位国家: 德国
• 项目类别: Major Research Instrumentation
• 财政年份: 2020
• 资助国家: 德国
• 起止时间: 2019-12-31 至 无数据
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/449737235?language=en
• 关键词: 3D; STED; Microscope

Investigations of small subcellular compartments has been significantly improved by the use of localisation microscopy methods. These methods allow to look into the structure and dynamic of molecular signalling complexes and are today standard microscopy tools in cell biology. The gain of localisation precision is about tenfold in comparison to confocal microscopy and reach the resolution of electron microscopy. The main advantage is the much simpler preparation of the probes and the possibility to use localisation microscopy also in living cells. Stochastic approach as Photoactivation localisation microscopy (PALM) or Stochastic Optical Reconstruction Microscopy (STORM) is most obvious in the gain of single fluorophore information, but relay on the acquisition of many pictures and precise reconstructions. The deterministic approach, called Stimulated Emission Depletion microscopy (STED) use a defined light pattern to scan the sample and allow to correlate much faster the localisation of molecules to each other. The optimization of detectors and illumination algorithms has improved the usability of STED microscopy in living cells. Implementation of adaptive optics or corrections in the optical components allow also a nearly similar localisation precision in all three spatial dimensions. With these improvements the STED microscopy is now relevant for a broad spectrum of samples. With the acquisition of a STED microscope we like to provide a valid tool for the growing number of molecular/cell biological working groups in the biology department of the Johannes Gutenberg University Mainz to conduct their research. The use of pulsed lasers within the STED microscope offers in addition the possibility to use the fluorescent live time of the fluorophores and investigate their interactions in relevant dimensions to their local arrangement.

小亚细胞区室的调查已显着改善使用定位显微镜方法。这些方法允许观察分子信号复合物的结构和动态，并且是当今细胞生物学中的标准显微镜工具。与共聚焦显微镜相比，定位精度提高了约10倍，达到了电子显微镜的分辨率。主要优点是探针的制备简单得多，并且有可能在活细胞中使用定位显微镜。随机方法如光活化定位显微术（PALM）或随机光学重建显微术（STORM）在获得单个荧光团信息方面最为明显，但依赖于许多图像的获取和精确的重建。被称为受激发射耗尽显微镜（STED）的确定性方法使用定义的光模式来扫描样品，并允许更快地将分子的定位相互关联。探测器和照明算法的优化提高了STED显微镜在活细胞中的可用性。自适应光学器件或光学部件中的校正的实现也允许在所有三个空间维度中的几乎类似的定位精度。随着这些改进，STED显微镜现在适用于广泛的样品。随着STED显微镜的收购，我们希望为美因茨约翰尼斯古滕贝格大学生物系越来越多的分子/细胞生物学工作组提供有效的工具来进行研究。在STED显微镜内使用脉冲激光提供了另外的可能性，使用荧光团的荧光寿命，并研究它们在相关维度上的相互作用。