Lightsheet-Microscopy-System

光片显微镜系统

• 批准号: 518201933
• 金额: --
• 依托单位: Otto-von-Guericke-Universität Magdeburg
• 依托单位国家: 德国
• 项目类别: Major Research Instrumentation
• 财政年份: 2023
• 资助国家: 德国
• 起止时间: 2022-12-31 至 无数据
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/518201933?language=en
• 关键词: Lightsheet; Microscopy; System

A white light laser Lightsheet microscope is proposed for the interdisciplinary research platform "Multiparametric Bioimaging and Cytometry" (MPBIC) of the Medical Faculty of the Otto-von-Guericke-Universität Magdeburg. The MPBIC platform is the central research infrastructure for light microscopy and flow cytometry approaches in the medical faculty within the research focus "Immunology and molecular medicine of inflammation". The imaging techniques currently available in the MPBIC (multiphoton-microscopy, confocal microscopy, wide-field microscopy) allow for the visualization of small samples or small sections of larger samples with high resolution. Larger samples have to be cut into thin sections due to the restricted penetration and emission depth of the exciting or the emitted light, which is leading to preparation artefacts and sample loss. However, various research objectives require the analyses of cell infiltrates or tissue maladaptation/structural alterations in the intact three-dimensional tissue context, which can be not addressed with thin sections (or only incompletely). The multi-photon intravital microscopy, which is a well-established procedure in the MPBIC platform, allows for the analysis of cell dynamics in the three-dimensional tissue but is limited to small sample regions despite the high penetration depth of the exciting laser. Consequently, this method is only limitedly useful for research projects addressing inflammatory processes, which are not homogeneously dispersed, but occurring in form of infiltration or infection spots. Recently, various methods of whole-mount staining and subsequent tissue clearing have been developed that enable the detection of structures in whole intact organs with fluorochrom-conjugated antibodies. Since, in practice, the combination of clearing with the scanning techniques available in the MPBIC leads to extremely long measuring times, this procedure is unsuitable for a broad application in the faculty. Therefore, the visualization and analysis of whole intact organs or tissues is not possible with the currently present microscopes or in a very restricted number. The herein proposed lightsheet-microscope will cover the increasing need in the faculty for fast 3D-analyses of large and intact biological samples (organs of experimental animals, organoids, and human biopsies) from the macro into the micro level. This will enable the detection of anatomical-structurally alterations/modifications in the tissue level up to infiltrations of immune cells, or structural alterations in inhomogeneously dispersed inflammation- or infection spots with cellular-subcellular resolution, which is indispensable for a number of promising running and planned research projects.

马格德堡奥托冯格里克大学医学院的跨学科研究平台“多参数生物成像和细胞计数”（MPBIC）提出了一种白色激光光片显微镜。MPBIC平台是研究重点“炎症的免疫学和分子医学”内医学系光学显微镜和流式细胞术方法的中心研究基础设施。MPBIC中目前可用的成像技术（多光子显微镜、共聚焦显微镜、宽视场显微镜）允许以高分辨率可视化小样品或较大样品的小切片。由于激发光或发射光的穿透和发射深度有限，较大的样品必须切割成薄片，这会导致制备伪影和样品损失。然而，各种研究目标需要在完整的三维组织背景下分析细胞浸润或组织适应不良/结构改变，这无法用薄切片（或仅不完全）解决。多光子活体显微镜是MPBIC平台中一种成熟的程序，可以分析三维组织中的细胞动力学，但仅限于小样本区域，尽管激发激光的穿透深度很高。因此，该方法仅有限地用于解决炎症过程的研究项目，其不是均匀分散的，而是以浸润或感染点的形式发生。最近，已经开发了各种各样的整体包埋染色和随后的组织清除方法，这些方法使得能够用荧光染料缀合的抗体检测完整器官中的结构。由于在实践中，MPBIC中可用的扫描技术与清除相结合导致极长的测量时间，因此该程序不适合在教师中广泛应用。因此，整个完整的器官或组织的可视化和分析是不可能的与目前的显微镜或在一个非常有限的数量。本文提出的光片显微镜将涵盖从宏观到微观层面对大型完整生物样品（实验动物器官，类器官和人类活检）进行快速3D分析的日益增长的需求。这将能够检测组织水平的解剖结构改变/修改，直到免疫细胞的浸润，或不均匀分散的炎症或感染点的结构改变，具有细胞亚细胞分辨率，这对于许多有前途的运行和计划的研究项目是必不可少的。