As part of the Leibniz ScienceCampus InfectoOptics we work in the project HoT-AiM (“High end optical technologies for the analysis of intracellular, membrane-influencing infection processes” ) in collaboration with other groups at the Beutenberg to characterize the dynamic membrane alterations in host cells mediated by the human pathogenic fungi Candida albicans und Aspergillus fumigatus. However, to tackle the problem and gain deeper insights of intracellular infections, the development of new imaging techniques is necessary.
holoSTORM is a new approach to use holographic patterns in combination with the STORM (SMLM) approach. This allows us to produce a ‘Region-of-Interest” (ROI)-Illumination in SMLM. In the context of entire cells the ROI illumination reduces -similar to confocal illumination- the background signal of out-of-focus planes in SMLM which normally depletes the image quality significantly. In addition, we want to use this approach to change the holographic pattern illumination adaptively to produce zero intensities at the assumed position of the fluorophores. This allows us to determine the localization of fluorophores with a nanometer precision with a highly reduced photon flux .