Confocal spinning disk high throughput screening microscope

• Subject Area: Medicine
• Term: Funded in 2021
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 459525245

We are applying for funding to acquire a confocal imaging cytometer that is urgently needed in several ongoing and planned interdisciplinary research projects.The possibilities, but also the technical requirements in high-end immunofluorescence microscopy have steadily expanded during the last few years and our current needs in this area cannot be met anymore with the available microscopy infrastructure on Charité Campus Mitte (one out of 3 Charité sites in Berlin). Recently, this has led to significant delays in our microscopy experiments in many projects and also affected the quality of the data we were able to generate. Furthermore, because of technical limitations of the available microscopes, we were not able to establish some more demanding experimental setups that would have greatly benefitted existing projects and are required to establish new research directions.Recently, enormous progress has been made in the design of microscopes and lots of new advanced applications in biology have emerged: more sensitive cameras, larger field of view and increasingly fully automated image acquisition workflows, as well as new optical designs make it possible to acquire high quality confocal images from hundreds of samples in 3D over long periods of time. Thus, it becomes possible to systematically track single cells in 3D over time in their natural environment, which is essential for a better understanding of many biological processes that are of immediate relevance in medicine: examples include the development of cancer, tracking of the immune response within tissues, but also many questions in cell biology that require for example experiments in organoids.Another important application for the imaging cytometer will be to obtain high-resolution confocal images of histological specimens from patients, fluorescently stained with new multiplexed protocols that enable sequential staining of the same slide for dozens of proteins. Acquisition of such images with the equipment currently available on campus is extremely time-consuming and therefore practically not feasible. The included image analysis package will also allow us to analyze the data on the fly during acquisition in an automated manner in high throughput.In summary, the confocal imaging cytometer will ensure the availability of cutting edge microscopy equipment for high-throughput 3D live-cell imaging coupled with dedicated software appropriate for quantification of such large amounts of data and help help us to establish new microscopy-based assays. It will also make it easier for members of the involved groups to get enough microscopy time for existing projects, which has become increasingly difficult relying on the existing equipment on campus. In the included application forms, we include a detailed plan how the cytometer will be used as benefit many existing and planned research projects of seven different labs that act as coapplicants.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Konfokales Spinning Disk Hochdurchsatz-Screening Mikroskop
• Instrumentation Group: 5042 Mikroskope für Hochdurchsatz und Screening
• Applicant Institution: Charité - Universitätsmedizin Berlin
• Leader: Professor Dr. David Horst