NANO-scale VIsualization to understand Bacterial virulence and invasiveness - based on fluorescence NANOscopy and VIBrational microscopy

ended

In an interdisciplinary project, we will prototype a next-generation super-resolution microscope (SRM) and demonstrate its capability to bring about a major leap forward in our understanding of inter- and intracellular processes, and thus the cellular origin of diseases. Based on the recently invented MINFLUX concept, which pushes spatial resolution an order of magnitude beyond any other SRM technique, and by concerted development of detector technologies, lasers and image acquisition procedures, we will be able to retrieve information, not within reach by any other photonics-based technique. By extending operation to the near infrared, a hitherto un-accessible spectral range for SRM, we will strongly reduce phototoxicity and scattering, increase penetration depth and provide an additional spectral window for multiplexing. The developed prototype will allow nanometer-scale protein localization patterns to be resolved and to be placed in a cellular context by overlaid morphological, biochemical and metabolic images generated by label-free stimulated Raman scattering (SRS) and two-photon excitation (TPE). In a lead application, we will use the unique capabilities of the to-be-developed technology to study the molecular mechanisms underlying pneumococcal disease, largely attributed to localization patterns of specific bacterial surface proteins, and their intricate interactions with immune and host target cells. Pneumococci are a major contributor to morbidity and mortality worldwide and we aim to provide vital information which can lead to new treatments and vaccines. We will also offer hands-on access to the technology to researchers from both academia and industry in an open demonstration facility. Together with the lead application, this will generate demand for microscopes, lasers and detectors, which the industrial partners will develop subsequent to this project based on the prototypes, further strengthening Europe´s industrial position in the microscopy field.