Advanced Confocal Laser-Scanning-Microscope

先进的共焦激光扫描显微镜

• 批准号: 503201387
• 金额: --
• 依托单位: Freie Universität Berlin
• 依托单位国家: 德国
• 项目类别: Major Research Instrumentation
• 财政年份: 2022
• 资助国家: 德国
• 起止时间: 2021-12-31 至 无数据
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/503201387?language=en
• 关键词: Advanced; Confocal; Laser; Scanning; Microscope

In this proposal we apply for funding of a state-of-the-art, fast and highly sensitive confocal laser scanning microscope (CLSM), with spatial (super)resolution beyond the diffraction limit and spectral detection. Both research units at the faculty Biology, Chemistry and Pharmacy and the integrated core unit BioSupraMol Optical Microscopy of Freie Universität Berlin will highly benefit from installation of this instrument as it will permit fast, but highly sensitive image acquisition relying on a high pixel dwell time for quantitative densitometric measurements in 3D and for high-throughput screening of biological samples and tissues, without affecting the signal-to-noise ratio. The novel detection technology will improve the lateral, and even more the axial resolution, permitting the detection of objects beyond the diffraction limit of classical confocal microscopes. The versatility of the instrument is reflected by a high scanning speed in combination with an improved signal-to-noise ratio and enhanced spatial resolution, so that fast in situ screenings of subcellular nano-structures (below 200 nm) even in large volumes are possible. The highly sensitive detector elements permit simultaneous spectral detection of more than ten fluorophores in the visible spectrum. The system relies on direct de-mixing of the different fluorophores in situ and therefore offers not only efficient multichannel imaging with a single scan, but also elimination of (chlorophyll-) autofluorescences. In addition, the novel system will provide the possibility to detect molecule interactions via Förster Resonance Energy Transfer (FRET), either by spectral FRET or by Flurorescence Lifetime Imaging (FLIM). Importantly, the system will bridge a substantial gap at the optical microscopy units at Freie Universität Berlin, as no other system currently combines high speed and superresolution with spectral demixing. The new microscope will be incorporated in the DFG funded Core Facility BioSupraMol and be located in the laboratories of the research building SupraFAB which is optimized for majors instrumentations.

在本提案中，我们申请资助最先进、快速、高灵敏度的共焦激光扫描显微镜（CLSM），其空间（超）分辨率超出了衍射极限和光谱检测。 、化学和制药以及柏林自由大学的集成核心单元 BioSupraMol 光学显微镜将极大地受益于该仪器的安装，因为它将允许依赖于高像素停留时间的快速但高灵敏度的图像采集3D 光密度测量以及生物样本和组织的高通量筛选，而不影响信噪比，新颖的检测技术将提高横向分辨率，甚至更高的轴向分辨率，从而能够检测超出范围的物体。该仪器的多功能性体现在高扫描速度、改进的信噪比和增强的空间分辨率上，从而可以快速原位筛选亚细胞纳米结构。即使是大体积（低于 200 nm），高灵敏度的检测器元件也可以同时对可见光谱中的十多个荧光团进行光谱检测。该系统依赖于不同荧光团的直接分离，因此不仅提供。通过单次扫描实现高效多通道成像，同时消除（叶绿素）自发荧光。此外，该新型系统将提供通过福斯特共振能量转移检测分子相互作用的可能性。 （FRET），无论是通过光谱 FRET 还是通过荧光寿命成像（FLIM） 重要的是，该系统将弥补柏林自由大学光学显微镜单位的巨大差距，因为目前没有其他系统将高速和超分辨率与光谱分离相结合。新显微镜将纳入 DFG 资助的核心设施 BioSupraMol 中，并位于 SupraFAB 研究大楼的实验室中，该大楼针对专业进行了优化仪器仪表。