Inverted camera-based fluorescence microscope for widefield microscopy

该提案的目的是获得一台荧光显微镜，以支持第三方资助的科学项目，研究形成病理生理条件的发育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。主要目标是描述和结合组织中细胞群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细胞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进展。另一个核心方面是确定在疾病期间组织微环境和细胞类型的代谢状态如何变化并形成疾病进展。荧光显微镜打算与现有的共聚焦显微镜结合使用，为科学项目带来几个积极的协同效应。这包括：1)在极短的时间内对大量样品进行分析；2)有针对性地、基于信息的选择样品中感兴趣的区域，以便后续使用共聚焦显微镜；3)缓解大量使用的共聚焦显微镜，以优化其特定技术的应用。为了实现这些协同作用，荧光显微镜必须在技术上配备，以最高的采集速度对样品进行精确测量，并与现有共聚焦显微镜高度互补，以确保在它们的联合应用中获得最大的效益。这需要能够以高分辨率和精度在不同的三维标本（如组织和类器官模型）中以高速度测量多个荧光参数。这些要求通过基于相机的宽视场显微镜的概念来满足，该显微镜可以去除杂散光。该显微镜将进一步研究快速的细胞内信号传导过程，并以高度加速的速度检测大型样品制备中的稀有细胞群。选择所需仪器的一个关键因素是它与现有共聚焦显微镜的互补性，这确保了两种仪器的即时组合，并在采集速度上获得最大增益。这将使科学项目的预期升级和扩展达到最佳状态。

The objective of this proposal is to acquire a fluorescence 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. The fluorescence microscope is intended to be used in combination with an existing confocal microscope, resulting in several positive synergistic effects for the scientific projects. These are 1) the analysis of a high number of samples in a drastically shortened time frame, 2) targeted, information-based selection of regions of interest in samples for the following use of the confocal microscope and 3) relief of the heavily used confocal microscope for optimal application of its specific technologies. To achieve these synergies, the fluorescence microscope has to be technically equipped to perform precise measurements in samples with highest acquisition speed, with a high degree of complementarity to the existing confocal microscope to ensure a maximal benefit in their combined application. This requires the ability to measure multiple fluorescence parameter with highly increased speed in diverse, three-dimensional specimen such as tissues and organoid models with high resolution and precision. These requirements are met by the concept of a camera-based widefield microscope that can remove stray light. This microscope will further allow the investigation of rapid intracellular signaling processes and the detection of rare cell populations in large sample preparations with highly accelerated speed. A key factor for the choice of the requested instrument was its complementarity to the existing confocal microscope, which ensures the instant combination of both instruments with a maximal gain in acquisition speed. This will enable the desired upscaling and extension of the scientific projects optimally.