Confocal-laser-scanning-microscope

• Subject Area: Microbiology, Virology and Immunology
• Term: Funded in 2024
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 544270579

Bacterial pathogens are currently a major threat worldwide, and the increasing loss of treatment options and lack of vaccination options will significantly exacerbate this threat in the future. In order to understand the mechanisms of disease development, a fundamental understanding of the interaction of pathogenic bacteria with human cells is important, especially for the development of diagnostics and new therapeutics. Intracellular bacterial pathogens are a particular challenge in this context, as they can evade the host's immune response and the effect of antimicrobial drugs by hiding in their host cells. Recently, organoids and organ-on-chip microfluidic technologies have been increasingly used as experimental systems to study host-pathogen interactions, enabling the study of infections similar to in vivo situations. This has greatly expanded our understanding of how infectious diseases might develop in vivo. Given the inherent three-dimensional (3D) architecture of such samples and the different time scales of infection-related processes (from minutes to days), there are specific requirements for microscopes that can be used to resolve the biological and infection-related processes in time and space. To date, we have used a Leica TCS SP5 confocal laser scanning microscope (LSM) for this work, for which the manufacturer no longer offers maintenance after 12 years of use due to a lack of spare parts and for which several laser light sources have already failed. We are therefore applying for a replacement confocal LSM for the visualization and quantification of 3D samples with living cells (infected or sterile) over short and long periods of time. The replacement acquisition is intended to create the prerequisites for continuing to develop and use state-of-the-art microscopy methods for research of infectious diseases. These include the time-resolved observation of infection processes on living objects, fluorescence resonance energy transfer (FRET) to visualize the interaction of host factors and bacterial factors and fluorescence recovery after photobleaching (FRAP) to observe dynamic processes in host cells in response to infection. These microscopy techniques are now internationally standard techniques for resolving dynamic processes at the molecular level. High-resolution techniques such as expansion microscopy are also used. The replacement microscope is to be installed in the S2-approved microscopy room of the Chair of Microbiology for the numerous projects in the field of host-pathogen interaction.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Konfokales Laserscanning-Mikroskop
• Instrumentation Group: 5090 Spezialmikroskope
• Applicant Institution: Julius-Maximilians-Universität Würzburg
• Leader: Professor Dr. Thomas Rudel