Advanced confocal laser scan system for imaging fixed and living biological samples

我们申请一种先进的激光共聚焦扫描显微镜，具有增强的激发和检测能力。应用显微镜对于提高汉堡大学医学中心的研究人员的成像质量和能力是必不可少的。英国皇家壳牌对共焦显微镜不断增长的需求现在已经供不应求。在过去的几年里，在一些关键的时间段，我们记录了极长的等待时间来预订共焦高分辨率成像切片，以及深夜和周末切片的大量使用。我们还经历了关键时刻，当时UKE显微镜成像设备(UMIF)的一个共焦显微镜因维修而不可用，因为这干扰了几篇高级论文的修改。事实上，大量使用的相继过时的显微镜发生故障的可能性越来越大，维修时间在一到三周之间。由于所有这些原因，在可用成像插槽没有强劲增加的情况下，UKE研究人员将忍不住寻找成像设施的替代方案，并可能自行启动主要的仪器建议。这将导致成像资源的次优分布。由于申请显微镜将用于当地的成像设施，其配置必须是通用的，并且整体设置应该是可升级的。这样，我们可以保证，在其整个生命周期内，这台显微镜将得到充分利用，因为它也可以用于本申请中未列出的成像项目。具体地说，这种显微镜应该能够产生固定的和活的样品的高分辨率图像，比透镜衍射的有限分辨率高2倍。重要的是，该成像平台还应提供成像速度和光学分辨率的最佳组合，以便能够可视化和跟踪活体样本中的亚细胞结构。这些要求可以通过在光学(物镜)、激发源(激光)和探测器(增强型PMT、GaAsP探测器以及先进探测器阵列)方面配备更好的技术解决方案的共焦显微镜来满足。最后，应用的显微镜必须是一个通用的平台，用于对广泛的固定和活的生物样本进行成像，包括免疫细胞(如巨噬细胞或小胶质细胞)、感染各种细菌的细胞(如伯氏疏螺旋体)、复杂模型(如小导管)、三维肿瘤模型或有机体，以及细胞内结构，如肌动蛋白细丝、微管、内质网或细胞器，如足体和吞噬体。

We apply for an advanced confocal laser scanning microscope with enhanced excitation and detection capabilities. The applied-for microscope will be essential for increasing the imaging quality and capacity available to the researchers working at the University Medical Center Hamburg. The constantly increasing demand for confocal microscopy at the UKE is now exceeding the supply. In the last years, in some critical time periods, we registered extremely long waiting times for booking confocal high resolution imaging sections and a massive usage of late night and weekend sections. We also experienced critical moments when one of the confocal microscopes of the UKE Microscopy Imaging Facility (UMIF) was not available due to repairs, as this interfered with the revision of several high-class papers. Indeed, failures are increasingly likely happen to heavily used successively obsolete microscopes, with repair times lasting between one and three weeks. Due to all these reasons, without a strong increase in available imaging slots, UKE researchers will be tempted to find alternatives to the imaging facility and probably start major instrumentation proposals on their own. This would result in a suboptimal distribution of the imaging resources. Since the applied-for microscope will be used in the local imaging facility, its configuration has to be versatile, and the overall set up should be upgradable. In this way, we can warrant that, for its entire life span, this microscope will be fully used, since it can also be employed for imaging projects that are not listed in this application. Specifically, this microscope should be able to produce high resolution pictures, up to 2 times better than lens diffraction limited resolution, of fixed and living samples. Importantly, this imaging platform should also provide an optimal combination of imaging speed and optical resolution in order to permit to visualize and follow sub-cellular structures in living samples. These requirements can be fulfilled by a confocal microscope that is equipped with better technological solutions in term of optics (objective lenses), excitation sources (lasers) and detectors (enhanced PMTs, GaAsP detectors as well as advanced detector arrays). Finally, the applied-for microscope has to be a versatile platform for imaging a wide range of fixed and living biological samples as, including immune cells such as macrophages or microglial cells, cells infected by a variety of bacteria such as Borrelia burgdorferi, complex models such as canaliculi, three dimensional tumour models, or organoids, and intracellular structures such as actin filaments, microtubules, endoplasmic reticulum, or organelles such as podosomes and phagosomes.