Screening microscope with confocal functionality

• Subject Area: Neurosciences
• Term: Funded in 2021
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 466742891

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.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Screening-Mikroskop mit Konfokalfunktion
• Instrumentation Group: 5042 Mikroskope für Hochdurchsatz und Screening
• Applicant Institution: Universität des Saarlandes
• Leader: Professor Dr. Markus Hoth