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

先进的共焦激光扫描系统，用于对固定和活体生物样本进行成像

• 批准号: 496964877
• 金额: --
• 依托单位: Universität Hamburg
• 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/496964877?language=en
• 关键词: Advanced; confocal; laser; scan; system

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.

我们申请了一个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。应用显微镜将是必不可少的，以提高成像质量和能力提供给研究人员在汉堡大学医学中心工作。UKE对共焦显微镜的需求不断增加，现在已经超过了供应。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预订共焦高分辨率成像部分和大量使用深夜和周末部分。我们还经历了关键时刻，当UKE显微成像设施（UMIF）的共焦显微镜之一由于维修而无法使用时，因为这干扰了几篇高级论文的修订。事实上，频繁使用的相继过时的显微镜发生故障的可能性越来越大，修复时间持续一到三周。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研究人员将试图找到成像设施的替代品，并可能自己开始主要的仪器建议。这将导致成像资源的次优分布。由于申请的显微镜将用于本地成像设施，其配置必须是通用的，整体设置应该是可拆卸的。通过这种方式，我们可以保证，在其整个寿命期间，该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。具体来说，这种显微镜应该能够产生高分辨率的图片，高达2倍以上的透镜衍射极限分辨率，固定和活的样品。重要的是，该成像平台还应该提供成像速度和光学分辨率的最佳组合，以便允许可视化和跟踪活样品中的亚细胞结构。这些要求可以通过共聚焦显微镜来满足，该显微镜在光学器件（物镜）、激发源（激光器）和检测器（增强型PMT、GaAsP检测器以及先进的检测器阵列）方面配备了更好的技术解决方案。最后，所应用的显微镜必须是用于对广泛的固定和活的生物样品成像的通用平台，所述生物样品包括免疫细胞如巨噬细胞或小胶质细胞、被多种细菌感染的细胞如伯氏疏螺旋体、复杂模型如小管、三维肿瘤模型或类器官，以及细胞内结构如肌动蛋白丝、微管、内质网、或细胞器如足体和吞噬体。