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

我们申请一种先进的共聚焦激光扫描显微镜，具有增强的激发和检测能力。申请的显微镜将是必不可少的，以提高成像质量和可用的研究人员在大学医学中心汉堡工作。在英国大学，对共聚焦显微镜不断增长的需求现在已经超过了供应。在过去的几年里，在一些关键时期，我们记录到预约共聚焦高分辨率成像区域的等待时间非常长，并且深夜和周末区域的大量使用。我们还经历了一个关键时刻，当UKE显微镜成像设施（UMIF）的一个共聚焦显微镜由于维修而无法使用，因为这干扰了几篇高级别论文的修改。的确，大量使用的过时显微镜出现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，如果没有强大的可用成像插槽的增加，UKE的研究人员将试图寻找成像设备的替代品，并可能开始自己的主要仪器建议。这将导致成像资源的次优分布。由于申请的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，在它的整个生命周期，这台显微镜将被充分利用，因为它也可以用于成像项目，没有在这个应用程序中列出。具体来说，这种显微镜应该能够产生高分辨率的图片，高达2倍的透镜衍射极限分辨率，固定和活的样品。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，以便在活样品中可视化和跟踪亚细胞结构。这些要求可以通过在光学（物镜），激发源（激光器）和探测器（增强型pmt， GaAsP探测器以及先进的探测器阵列）方面配备更好的技术解决方案的共聚焦显微镜来满足。最后，申请的显微镜必须是一个多功能的平台，用于成像广泛的固定和活的生物样品，包括免疫细胞，如巨噬细胞或小胶质细胞，被各种细菌感染的细胞，如伯氏疏疏体，复杂模型，如小管，三维肿瘤模型，或类器官，细胞内结构，如肌动蛋白丝，微管，内质网，或细胞器，如足小体和吞噬体。

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.