Inverted camera-based fluorescence microscope for widefield microscopy

• Subject Area: Microbiology, Virology and Immunology
• Term: Funded in 2020
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 442405234

The objective of this proposal is to acquire a fluorescence 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. The fluorescence microscope is intended to be used in combination with an existing confocal microscope, resulting in several positive synergistic effects for the scientific projects. These are 1) the analysis of a high number of samples in a drastically shortened time frame, 2) targeted, information-based selection of regions of interest in samples for the following use of the confocal microscope and 3) relief of the heavily used confocal microscope for optimal application of its specific technologies. To achieve these synergies, the fluorescence microscope has to be technically equipped to perform precise measurements in samples with highest acquisition speed, with a high degree of complementarity to the existing confocal microscope to ensure a maximal benefit in their combined application. This requires the ability to measure multiple fluorescence parameter with highly increased speed in diverse, three-dimensional specimen such as tissues and organoid models with high resolution and precision. These requirements are met by the concept of a camera-based widefield microscope that can remove stray light. This microscope will further allow the investigation of rapid intracellular signaling processes and the detection of rare cell populations in large sample preparations with highly accelerated speed. A key factor for the choice of the requested instrument was its complementarity to the existing confocal microscope, which ensures the instant combination of both instruments with a maximal gain in acquisition speed. This will enable the desired upscaling and extension of the scientific projects optimally.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Inverses kamerabasiertes Weitfeld-Fluoreszenzmikroskop
• Instrumentation Group: 5000 Labormikroskope
• Applicant Institution: Technische Universität München (TUM)
• Leader: Dr. Jan Philipp Böttcher