Confocal laser scanning microscope with FLIM (Teilfinanzerung)

• Subject Area: Basic Research in Biology and Medicine
• Term: Funded in 2025
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 566642683

Scientists at Institute of Biochemistry II (IBC2) and their partner institutions at Goethe University Frankfurt, Medical Faculty are investigating fundamental molecular processes in the fields of biochemistry, cell biology, and biomedicine. We aim i) to uncover and characterize novel aspects of the regulation of selective organellophagy and the functional interplay between different organelles, and ii) to dissect cellular responses to different forms of stress, such as pathogen invasion, proteotoxic or genotoxic insults, nutrient starvation, and inflammation. Image-based approaches, especially fluorescence confocal laser scanning microscopy, are instrumental for these studies. However, the currently accessible instrumentation only allows fluorescence- intensity-based measurements, mostly in fixed cells, with a limited choice of lasers and, hence, fluorophores that can be utilized. Neither high-resolution live cell imaging nor functional imaging can be performed. Studying dynamic processes, such as formation of organelle contacts, post-translational modification of proteins and organelles, lipid composition of membranes, protein-protein or protein-DNA/RNA interactions, protein trafficking, and formation and composition of nuclear condensates, requires rapid and gentle imaging in live cells with enhanced resolution, allowing sample analysis for a prolonged period of time with low phototoxicity during image acquisition. This is currently not possible. To overcome these limitations, a new fluorescence confocal laser scanning microscope with a fully integrated fluorescence lifetime imaging (FLIM) module shall be purchased. The implementation of FLIM will provide new insights into molecular interactions and cellular dynamics that cannot be studied using the present instrumentation. In contrast to measuring the intensity of light emitted by an excited fluorophore returning to its ground state, FLIM records the average time that the fluorophore remains in the excited state, which is a unique property of each fluorophore. Lifetime information is influenced by the environment, for example, by binding to other molecules. Therefore, FLIM is a powerful tool for investigating molecular functions, interactions, and the environment, especially when combined with Förster resonance energy transfer (FRET). FLIM is the core requirement of the sought-after laser scanning microscope. Together with a white light laser (WLL), simultaneous imaging of new fluorophore combinations would become possible and would allow extraction of information from confocal images coming close to super-resolution territory. To carry out the ambitious studies proposed, a fluorescence confocal laser scanning microscope with a fully integrated FLIM module is needed. It will provide exceptional image quality, versatility, ease of use, and advanced data analysis capabilities, and facilitate a deeper understanding of cellular processes by revealing information that is currently inaccessible.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Konfokales Laser-Scanning-Mikroskop mit FLIM (Teilfinanzierung)
• Instrumentation Group: 5090 Spezialmikroskope
• Applicant Institution: Goethe-Universität Frankfurt am Main
• Leader: Dr. Anja Bremm