Confocal Laser Scanning Microscope with Spinning Disk Technology

采用转盘技术的共焦激光扫描显微镜

• 批准号: 522417173
• 金额: --
• 依托单位: Friedrich-Alexander-Universität Erlangen-Nürnberg
• 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/522417173?language=en
• 关键词: Confocal; Laser; Scanning; Microscope; Spinning

OICE is a centre of excellence (central institute) of FAU and represents the centralised platform for high and ultra-high resolution imaging using light-based technologies for, among other things, living cells and tissues, organoids, intravital imaging and imaging of biological materials and membranes. OICE combines a Core Facility Unit (CFU), Exploratory Research Unit (ERU) and Educational Training Unit (ETU). The interaction of physics/engineering on the development side, with researchers from natural sciences, medicine and material sciences on the application side, and the well-founded training strengthens Erlangen as a research location and has established OICE nationally and also internationally as a recognised institute for light microscopy. The novel combination of real-time super resolution with spinning disc laser scanning allows for the first time long-term measurements of living three-dimensional samples with improved lateral resolution and, in combination with improved silicone immersion objectives, higher penetration depths with lower artefacts of > 100 µm. However, the speed and significantly lower illumination intensity of an SD-LSM is retained. Thus, this Super Resolution SD-LSM for long-term live cell observation replaces the previous SD-LSM (acquired in 2013) and complements the existing devices such as the multiphoton (intravital imaging) LSM and light sheet microscopy (fixed samples). At the OICE, there is still a high demand for spinning disc technology, especially considering the technological advancements in resolution, penetration depth and better camera detectors. The majority of the > 30 research groups/year that successfully use this technology use organoid models, live cell cultures or tissues to perform long-term spatio-temporal multichannel imaging (from several hours to whole days). The highest possible lateral (super) resolution for dynamic intracellular or membrane-associated processes is essential. The high thickness of the samples (organoids / tissue) requires a high penetration depth with minimised scattering to enable meaningful three-dimensional reconstruction and measurements, also over time. A large optical field of view that is as homogeneously illuminated as possible also minimises tile imaging with downstream stitching, which is associated with losses. In the medium term, this device should also relieve and replace the classic SD-LSM, which has now been in continuous operation for ten years (> 2,600 hours/year).

OICE是FAU的卓越中心（中央研究所），代表了使用基于光的技术进行高和超高分辨率成像的集中平台，用于活细胞和组织，类器官，活体成像以及生物材料和膜的成像。OICE由核心设施单位（CFU），探索性研究单位（ERU）和教育培训单位（ETU）组成。物理学/工程学在开发方面的相互作用，与自然科学，医学和材料科学的研究人员在应用方面的相互作用，以及有根据的培训加强了埃尔兰根作为研究地点的地位，并在国内和国际上建立了OICE作为公认的光学显微镜研究所。实时超分辨率与旋转圆盘激光扫描的新颖组合首次实现了对活体三维样品的长期测量，提高了横向分辨率，并与改进的硅胶浸没物镜相结合，实现了更高的穿透深度和更低的> 100 µm伪影。然而，SD-LSM的速度和显著较低的照明强度被保留。因此，这款用于长期活细胞观察的超分辨率SD-LSM取代了之前的SD-LSM（2013年获得），并补充了现有的设备，如多光子（活体成像）LSM和光片显微镜（固定样品）。在OICE，对旋转光盘技术的需求仍然很高，特别是考虑到分辨率、穿透深度和更好的相机探测器的技术进步。每年有超过30个研究小组成功使用这项技术，其中大多数使用类器官模型、活细胞培养物或组织进行长期时空多通道成像（从几小时到几天）。动态细胞内或膜相关过程的最高可能的横向（超）分辨率是必不可少的。样品（类器官/组织）的高厚度需要具有最小化散射的高穿透深度，以实现有意义的三维重建和测量，也随着时间的推移。尽可能均匀照明的大光学视场还使具有下游拼接的瓦片成像最小化，这与损耗相关联。从中期来看，该设备还将缓解并取代经典的SD-LSM，该设备现已连续运行十年（> 2，600小时/年）。