Confocal Laserscanning Microscope with fluorescence lifetime applications

• Subject Area: Basic Research in Biology and Medicine
• Term: Funded in 2025
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 559327492

The technique of confocal fluorescence microscopy represents a versatile approach in many aspects of scientific research, to specifically elucidate processes and structures in biological and medical samples or in the field of material science. The confocal optical sectioning allows for high contrast of single focal plane imaging or 3D analysis. Specific spectral separation of different signals is key in every fluorescent experiment, but especially, if many targets are combined in a single measurement, which is often the case in multiplexing approaches of medical experiments. Finetuning and iterative optimization of specific combinations of fluorescent dyes is crucial to avoid misleading crosstalk and to acquire high quality data. The process of spectral separation can further be supported by adding information about fluorescence lifetime. This data give insight into microenvironmental characteristics, specificity of a different signal with the same spectral properties, reduction of unspecific background signals or even interaction of different proteins in vivo. The latter can be achieved by observing an effect called Foerster resonance energy transfer (FRET), and is an important readout in e. g. photobleaching experiments (FRET-APB), or when determining the fluorescent lifetime of one of the fluorescent proteins fused to the proteins of interest (FRET-FLIM). Information like this is absolutely valuable for many life-science researchers, as it provides insights into protein-protein interaction at specific locations in vivo with high spatial resolution. Nevertheless, acquisition and analysis of dynamic processes is equally important to estimate dynamics of molecules or structures in timeseries experiments like fluorescence recovery after photobleaching (FRAP), if necessary, even in a sub-second range. Here, we apply for a confocal laser scanning microscope equipped with additional features for measuring fluorescence lifetime. The system applied for will give various institutes access to state-of-the-art instrumentation under supervision of a well experienced and trained team of our core facility, in order to increase the efficiency and quality of related research projects, while reducing the workload of existing instrumentation.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Konfokales Laserscanning Mikroskop mit Fluoreszenzlebenszeitanwendungen
• Instrumentation Group: 5090 Spezialmikroskope
• Applicant Institution: Heinrich-Heine-Universität Düsseldorf
• Leader: Professorin Dr. Stefanie Weidtkamp-Peters