Confocal Laser Scanning Microscope with Spinning Disk Technology

OICE是FAU的卓越中心(中央研究所)，代表着利用基于光的技术进行高分辨率和超高分辨率成像的中央平台，除其他外，用于活细胞和组织、有机体、生物体内成像和生物材料和膜的成像。OICE将核心设施股(CFU)、探索性研究股(ERU)和教育培训股(ETU)合并在一起。物理/工程在开发方面的互动，以及自然科学、医学和材料科学的研究人员在应用方面的互动，以及有根有据的培训加强了Erlangen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组合首次允许对活体三维样品进行长期测量，具有更高的横向分辨率，结合改进的硅胶浸泡物镜，可以获得更高的穿透深度和更低的&gt；100微米伪影。但仍保留了SD-LSM的速度和显著较低的照明强度。因此，这种用于长期活细胞观察的超分辨率SD-LSM取代了以前的SD-LSM(2013年收购)，并补充了现有的设备，如多光子(生命内成像)LSM和光片显微镜(固定样本)。在OICE，对旋转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方面的技术进步。每年成功使用这项技术的30个研究小组中的大多数使用有机模型、活细胞培养或组织来执行长期的时空多通道成像(从几个小时到一整天)。对于动态的细胞内或膜相关过程，尽可能高的横向(超)分辨率是必不可少的。高厚度的样品(有机物/组织)需要高渗透深度和最小的散射，以便能够进行有意义的三维重建和测量，也是随着时间的推移。尽可能均匀照明的大光学视场还可以最大限度地减少带有下游拼接的瓷砖成像，这与损失有关。从中期来看，该设备还将取代传统的SD-LSM，后者现在已经连续运行了10年(&gt；2600小时/年)。

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).