Confocal laser-scanning-microscope

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The Faculty of Biology and Biotechnology, specifically all groups working on plant model organisms, urgently need a confocal laser scanning microscope with high resolution and sensitivity. Access to confocal microscopy is very limited for the groups working on plants and to meet the increased demand for subcellular localization of proteins, live cell imaging of dynamic structures such as autophagosomes, vesicles and other small structures (e.g., processing bodies, amphisomes, Golgi etc.) and protein-protein interaction studies (FRET-FLIM) in living plant tissues, a state-of-the art confocal microscope is required. All proposed project aim (i) to localize or co-localize fluorescently tagged proteins under different conditions, and (ii) to perform protein-protein interaction studies using FRET-FLIM and/or BiFC. Projects from the Grefen, Ebert, Krämer, González Fuente and Üstün groups will also monitor dynamics of candidate proteins in different contexts. Moreover, the research groups of Ute Krämer, Suayb Üstün and Manuel González Fuente will perform FRAP experiments in their proposed projects. To guarantee the success of all proposed projects, it is vital to have access to a state-of-the art confocal microscope. The proposed device is characterized (i) by unique possibilities for the elimination of autofluorescence, which are of great importance for plant research, (ii) through high resolution and sensitivity and (iii) by using up to 8 simultaneous independent laser lines which allows the detection of multiple fluorophores in a sample. With the resonant scanner this microscope can scan images at high speed, comparable to that of a spinning disc confocal, which is essential to monitor highly mobile structures as well as detect weakly expressed fluorescent fusion proteins that are prone to bleaching. The integrated FLIM unit is unique for this device, simplifying the pre- and post-measurement data analysis. Using the FLIM unit will allow us to investigate fast processes in living samples (e.g., vesicle sorting/transport, autophagosome fusion events, signal transduction and protein-protein interactions).