Confocal Scanning Microscope with spectral detection and FLIM

具有光谱检测和 FLIM 功能的共焦扫描显微镜

• 批准号: 424926990
• 金额: --
• 依托单位: Technische Universität München (TUM)
• 依托单位国家: 德国
• 项目类别: Major Research Instrumentation
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 无数据
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/424926990?language=en
• 关键词: Confocal; Scanning; Microscope; spectral; detection

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.

该提案的目的是获得一台共聚焦显微镜，以支持第三方资助的科学项目，调查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、心血管和代谢疾病。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，以开发靶向治疗方法。主要目标是表征和联合收割机有关组织中细胞群空间定位及其功能状态的信息，并进一步详细研究移动的免疫细胞与实质或肿瘤细胞之间的动态相互作用，以了解细胞间通讯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和细胞类型中的代谢状态在疾病期间如何变化并形成疾病进展。关于组织中细胞群定位的详细信息只能通过高分辨率共聚焦显微镜获得，该显微镜在技术上配备有同时测量具有高精度的细胞类型特异性参数的许多组合。多个参数的这种测量的重要标准是在荧光团的激发和发射光的收集中的最大效率，与高灵敏度、低噪声光电探测器相结合。概述的项目在不同的体内系统，类器官模型和组织制备中进行，这些系统通常具有很强的自发荧光。因此，至关重要的是，研究仪器可以通过光谱分析和荧光寿命采集等技术来识别自发荧光。通过检测荧光寿命来表征自发荧光进一步提供了对组织和细胞的代谢状态进行新发现的巨大可能性，因为自发荧光根据代谢状态而变化。为了在活细胞的动态相互作用过程中以高分辨率研究快速过程，仪器必须配备相应的高速扫描头和高灵敏度检测器。光学信息的快速和高灵敏度采集进一步确保了此类分析所需的低水平光毒性。为了在细胞相互作用期间精确可视化精细亚细胞结构，还必须在超分辨率水平上进行成像-最佳地在分析参数的选择上没有任何限制。选择所需仪器的一个关键因素是不同技术的高标准模块化集成，确保以最佳方式组合应用这些技术的必要灵活性。