Screening microscope with confocal functionality

我们正在申请一种具有共聚焦功能的筛选显微镜。在生理细胞/组织培养条件下，最多可同时分析96（384）个平行条件，具有非常好的时间分辨率。在完整的96（384）孔中可以分辨出单细胞以及亚细胞结构，具有非常好的共聚焦分辨率。在测量过程中，所有物镜（包括高数值孔径的水浸物镜）都可以完全自动改变，从而将筛选实验与（自动）选定区域的高分辨率共聚焦测量相结合。并行测量不仅具有巨大的时间优势，而且还可以优化利用来自癌症患者的材料。此外，通过对一只动物的细胞/组织进行并行测量，3R原则（替换，减少，精炼）在动物实验研究中得到了很好的实施。特别是，人或鼠的免疫细胞和癌细胞的相互作用的分析将在我们的部门进行。对于这些实验，需要在96井（或384井）测量的同时获取共聚焦数据。对平行96孔（或384孔）实验的需求源于这样一个事实，即许多用献血者或癌症患者的免疫细胞进行的8小时（甚至24-48小时）实验不能连续进行，因为1。50种不同的条件下，8小时的实验大约需要20天，2)细胞不会持续那么长时间，3)测量结果将不再具有可比性。本资助的所有申请人必须对原始人类和/或小鼠单细胞或类器官进行平行测量，最好在数小时至数天内进行测量。同时，亚细胞结构必须在相同的测量分析，以了解分子过程。有了要求的具有共聚焦功能的筛选显微镜，亚细胞荧光测量和3D重建堆叠图像的分辨率小于200nm是可能的。因此，具有共聚焦功能的筛选显微镜非常适合执行这里描述的项目。它相对容易使用，因此可以预期，来自Homburg和saarbr<e:1> cken的许多团体将使用该仪器。通常，对大量数据的分析是一个主要问题。作为2017 - 2022年BMBF项目的一部分，我们可以为所有申请人提供半自动分析。为了在未来尽可能地自动化这些半自动分析，我们刚刚聘请了一名全职生物信息学家。

We are applying for a screening microscope with confocal functionality. Up to 96 (384) parallel conditions can be analyzed simultaneously with very good time resolution under physiological cell/tissue culture conditions. Single cells in the complete 96 (384) well can be resolved as well as subcellular structures with very good confocal resolution. All objectives (including the water immersion objectives with high numerical aperture) can be changed fully automatically during the measurement, so that screening experiments can be combined with high-resolution confocal measurements of (automatically) selected areas. Parallelized measurements not only have enormous time advantages, but also make it possible to make optimal use of the material from - in our case - cancer patients. Furthermore, the 3R principle (Replace, Reduce, Refine) is very well implemented in animal experimental research by parallelized measurements with cells/tissues of one animal.In particular, analyses of the interactions of human or murine immune and cancer cells will be performed in our department. For these experiments, it is necessary to measure in 96-well (or 384-well) while simultaneously acquiring confocal data. The need for parallel 96-well (or 384-well) experiments arises from the fact that many of the 8 (or even 24-48) hour-long experiments with immune cells from blood donors or cancer patients cannot be performed consecutively because 1.) 50 different conditions would take approximately 20 days for 8-hour experiments, 2.) the cells would not last that long, and 3.) the measurements would no longer be comparable. All applicants of this grant must perform parallelized measurements of primary human and/or murine single cells or organoids, preferably measured over hours to days. At the same time, subcellular structures must be analyzed in the same measurement to understand molecular processes. With the requested screening microscope with confocal functionality, subcellular fluorescence measurements and 3D reconstructions from stacked images with an xy resolution of less than 200 nm are possible.The screening microscope with confocal function is thus ideally suited to perform the projects described here. It is relatively easy to use, so it is to be expected that the instrument will be used by very many groups from Homburg and Saarbrücken. Often the analysis of the very large amount of data is a major problem. As part of a BMBF project funded from 2017 - 2022, we can support all applicants here with semi-automatic analyses. In order to automate these semi-automatic analyses as best as possible in the future, we have just a hired full-time bioinformatician.