Confocal Scanning Microscope with spectral detection and FLIM

• Subject Area: Microbiology, Virology and Immunology
• Term: Funded in 2019
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 424926990

The objective of this proposal is to acquire a confocal microscope for the support of third-party funded scientific projects investigating the molecular and cellular mechanisms that shape development and regulation of pathophysiological conditions. These conditions include cancer, chronic viral hepatitis, cardiovascular and metabolic diseases. The common aim of the projects is to achieve a better understanding of the complex dynamics and molecular mechanisms of multicellular processes within the affected organs and tissues, with the goal to develop targeted therapies.Main objectives are to characterize and combine information on the spatial localization of cell populations in tissues with their functional states, and further a detailed investigation of the dynamic interactions between mobile immune cells and parenchymal or tumor cells, in order to understand how intercellular communication determines disease initiation and progression. Another central aspect is to identify how metabolic states in tissue microenvironments and cell types change during disease and shape disease progression. Detailed information on the localization of cell populations in tissues can only be acquired with a high resolution confocal microscope, technically equipped to simultaneously measure many combinations of cell type-specific parameters with high accuracy. Important criteria for such measurements of multiple parameters are maximal efficiency in the excitation of fluorophores and in the collection of emitted light, combined with highly sensitive, low noise photodetectors. The outlined projects are pursued in different in vivo systems, organoid models, and tissue preparations, which often possess strong autofluorescence. It is therefore vital that the research instrument can identify autofluorescences through techniques such as spectral analysis and fluorescence lifetime acqusition. The characterization of autofluorescences via the detection of fluorescence lifetimes further provides the great possibility to make new discoveries for the metabolic states of tissues and cells, as autofluorescences change depending on metabolic states. In order to investigate fast processes with high resolution during the dynamic interaction of live cells, it is mandatory that the instrument is equipped with the corresponding high-speed scan head and highly sensitive detectors. Fast and highly sensitive acquisition of optical information further ensure the low level of phototoxicity needed for such analyses. For the accurate visualization of fine subcellular structures during the interaction of cells, it is further essential to perform imaging on the superresolution level – optimally without any restrictions in the choice of analyzed parameters. A key factor for the choice of the requested instrument was the high standard of modular integration of the different technologies, guaranteeing the necessary flexibility for the combined application of these technologies in an optimal way.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Konfokales Laser Scanning Mikroskop mit Spektraler Detektion und FLIM
• Instrumentation Group: 5090 Spezialmikroskope
• Applicant Institution: Technische Universität München (TUM)
• Leader: Dr. Jan Philipp Böttcher