TY  - JOUR
A1  - Trinks, Nora
A1  - Reinhard, Sebastian
A1  - Drobny, Matthias
A1  - Heilig, Linda
A1  - Löffler, Jürgen
A1  - Sauer, Markus
A1  - Terpitz, Ulrich
T1  - Subdiffraction-resolution fluorescence imaging of immunological synapse formation between NK cells and A. fumigatus by expansion microscopy
JF  - Communications Biology
N2  - 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.
KW  - biological fluorescence
KW  - fluorescence imaging
KW  - imaging the immune system
KW  - infectious diseases
KW  - super-resolution microscopy
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-264996
VL  - 4
IS  - 1
ER  - 
TY  - JOUR
A1  - Klamp, Tobias
A1  - Camps, Marta
A1  - Nieto, Benjamin
A1  - Guasch, Francesc
A1  - Ranasinghe, Rohan T.
A1  - Wiedemann, Jens
A1  - Petrášek, Zdeněk
A1  - Schwille, Petra
A1  - Klenerman, David
A1  - Sauer, Markus
T1  - Highly Rapid Amplification-Free and Quantitative DNA Imaging Assay
JF  - Scientific Reports
N2  - 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.
KW  - laboratory techniques and procedures
KW  - diseases
KW  - infectious diseases
KW  - assay systems
Y1  - 2013
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-130500
VL  - 3
IS  - 1852
ER  -