Confocal Scanning Microscope with spectral detection and FLIM

这项提议的目的是获得一种共聚焦显微镜，用于支持第三方资助的科学项目，该项目研究形成病理生理条件的发展和调节的分子和细胞机制。这些疾病包括癌症、慢性病毒性肝炎、心血管疾病和代谢性疾病。这些项目的共同目标是更好地了解受影响器官和组织内多细胞过程的复杂动力学和分子机制，目标是开发有针对性的治疗方法。主要目标是表征和结合有关组织中细胞群体的空间定位及其功能状态的信息，并进一步详细研究移动免疫细胞与实质或肿瘤细胞之间的动态相互作用，以了解细胞间通信如何决定疾病的发生和发展。另一个核心方面是确定组织微环境中的代谢状态和细胞类型在疾病过程中如何变化，并影响疾病的进展。有关细胞群体在组织中定位的详细信息只能通过高分辨率共聚焦显微镜获得，该显微镜的技术装备可以高精度地同时测量多种细胞类型特定参数的组合。多参数测量的重要标准是结合高灵敏度、低噪声的光电探测器，最大限度地提高荧光团的激发效率和发射光的收集效率。概述的项目在不同的活体系统、有机模型和组织制剂中进行，这些系统通常具有强烈的自体荧光。因此，研究仪器能够通过光谱分析和荧光寿命获取等技术识别自发荧光是至关重要的。通过检测荧光寿命来表征自体荧光，进一步为发现组织和细胞的代谢状态提供了极大的可能性，因为自体荧光的变化依赖于代谢状态。为了研究活细胞动态相互作用过程中高分辨率的快速过程，仪器必须配备相应的高速扫描头和高灵敏度的探测器。对光学信息的快速和高度敏感的获取进一步确保了此类分析所需的低水平的光毒性。为了准确显示细胞相互作用过程中的细微亚细胞结构，在超分辨率水平上进行成像是非常必要的--在分析参数的选择上不受任何限制。选择所要求的文书的一个关键因素是不同技术的模块集成的高标准，从而保证以最佳方式组合应用这些技术所需的灵活性。

The objective of this proposal is to acquire a confocal microscope for the support of third-party funded scientific projects investigating the molecular and cellular mechanisms that shape development and regulation of pathophysiological conditions. These conditions include cancer, chronic viral hepatitis, cardiovascular and metabolic diseases. The common aim of the projects is to achieve a better understanding of the complex dynamics and molecular mechanisms of multicellular processes within the affected organs and tissues, with the goal to develop targeted therapies.Main objectives are to characterize and combine information on the spatial localization of cell populations in tissues with their functional states, and further a detailed investigation of the dynamic interactions between mobile immune cells and parenchymal or tumor cells, in order to understand how intercellular communication determines disease initiation and progression. Another central aspect is to identify how metabolic states in tissue microenvironments and cell types change during disease and shape disease progression. Detailed information on the localization of cell populations in tissues can only be acquired with a high resolution confocal microscope, technically equipped to simultaneously measure many combinations of cell type-specific parameters with high accuracy. Important criteria for such measurements of multiple parameters are maximal efficiency in the excitation of fluorophores and in the collection of emitted light, combined with highly sensitive, low noise photodetectors. The outlined projects are pursued in different in vivo systems, organoid models, and tissue preparations, which often possess strong autofluorescence. It is therefore vital that the research instrument can identify autofluorescences through techniques such as spectral analysis and fluorescence lifetime acqusition. The characterization of autofluorescences via the detection of fluorescence lifetimes further provides the great possibility to make new discoveries for the metabolic states of tissues and cells, as autofluorescences change depending on metabolic states. In order to investigate fast processes with high resolution during the dynamic interaction of live cells, it is mandatory that the instrument is equipped with the corresponding high-speed scan head and highly sensitive detectors. Fast and highly sensitive acquisition of optical information further ensure the low level of phototoxicity needed for such analyses. For the accurate visualization of fine subcellular structures during the interaction of cells, it is further essential to perform imaging on the superresolution level – optimally without any restrictions in the choice of analyzed parameters. A key factor for the choice of the requested instrument was the high standard of modular integration of the different technologies, guaranteeing the necessary flexibility for the combined application of these technologies in an optimal way.