@article{TrinksReinhardDrobnyetal.2021, author = {Trinks, Nora and Reinhard, Sebastian and Drobny, Matthias and Heilig, Linda and L{\"o}ffler, J{\"u}rgen and Sauer, Markus and Terpitz, Ulrich}, title = {Subdiffraction-resolution fluorescence imaging of immunological synapse formation between NK cells and A. fumigatus by expansion microscopy}, series = {Communications Biology}, volume = {4}, journal = {Communications Biology}, number = {1}, doi = {10.1038/s42003-021-02669-y}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-264996}, year = {2021}, abstract = {Expansion microscopy (ExM) enables super-resolution fluorescence imaging on standard microscopes by physical expansion of the sample. However, the investigation of interactions between different organisms such as mammalian and fungal cells by ExM remains challenging because different cell types require different expansion protocols to ensure identical, ideally isotropic expansion of both partners. Here, we introduce an ExM method that enables super-resolved visualization of the interaction between NK cells and Aspergillus fumigatus hyphae. 4-fold expansion in combination with confocal fluorescence imaging allows us to resolve details of cytoskeleton rearrangement as well as NK cells' lytic granules triggered by contact with an RFP-expressing A. fumigatus strain. In particular, subdiffraction-resolution images show polarized degranulation upon contact formation and the presence of LAMP1 surrounding perforin at the NK cell-surface post degranulation. Our data demonstrate that optimized ExM protocols enable the investigation of immunological synapse formation between two different species with so far unmatched spatial resolution.}, language = {en} } @article{KlampCampsNietoetal.2013, author = {Klamp, Tobias and Camps, Marta and Nieto, Benjamin and Guasch, Francesc and Ranasinghe, Rohan T. and Wiedemann, Jens and Petr{\´a}šek, Zdeněk and Schwille, Petra and Klenerman, David and Sauer, Markus}, title = {Highly Rapid Amplification-Free and Quantitative DNA Imaging Assay}, series = {Scientific Reports}, volume = {3}, journal = {Scientific Reports}, number = {1852}, doi = {10.1038/srep01852}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-130500}, year = {2013}, abstract = {There is an urgent need for rapid and highly sensitive detection of pathogen-derivedDNAin a point-of-care (POC) device for diagnostics in hospitals and clinics. This device needs to work in a 'sample-in-result-out' mode with minimum number of steps so that it can be completely integrated into a cheap and simple instrument. We have developed a method that directly detects unamplified DNA, and demonstrate its sensitivity on realistically sized 5 kbp targetDNA fragments of Micrococcus luteus in small sample volumes of 20 mL. The assay consists of capturing and accumulating of target DNA on magnetic beads with specific capture oligonucleotides, hybridization of complementary fluorescently labeled detection oligonucleotides, and fluorescence imaging on a miniaturized wide-field fluorescence microscope. Our simple method delivers results in less than 20 minutes with a limit of detection (LOD) of,5 pMand a linear detection range spanning three orders of magnitude.}, language = {en} }