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  - Götz, Ralph
A1  - Panzer, Sabine
A1  - Trinks, Nora
A1  - Eilts, Janna
A1  - Wagener, Johannes
A1  - Turrà, David
A1  - Di Pietro, Antonio
A1  - Sauer, Markus
A1  - Terpitz, Ulrich
T1  - Expansion Microscopy for Cell Biology Analysis in Fungi
JF  - Frontiers in Microbiology
N2  - Super-resolution microscopy has evolved as a powerful method for subdiffraction-resolution fluorescence imaging of cells and cellular organelles, but requires sophisticated and expensive installations. Expansion microscopy (ExM), which is based on the physical expansion of the cellular structure of interest, provides a cheap alternative to bypass the diffraction limit and enable super-resolution imaging on a conventional fluorescence microscope. While ExM has shown impressive results for the magnified visualization of proteins and RNAs in cells and tissues, it has not yet been applied in fungi, mainly due to their complex cell wall. Here we developed a method that enables reliable isotropic expansion of ascomycetes and basidiomycetes upon treatment with cell wall degrading enzymes. Confocal laser scanning microscopy (CLSM) and structured illumination microscopy (SIM) images of 4.5-fold expanded sporidia of Ustilago maydis expressing fluorescent fungal rhodopsins and hyphae of Fusarium oxysporum or Aspergillus fumigatus expressing either histone H1-mCherry together with Lifeact-sGFP or mRFP targeted to mitochondria, revealed details of subcellular structures with an estimated spatial resolution of around 30 nm. ExM is thus well suited for cell biology studies in fungi on conventional fluorescence microscopes.
KW  - Expansion microscopy
KW  - fluorescence microscopy
KW  - fungi
KW  - sporidia
KW  - hyphae
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-202569
SN  - 1664-302X
VL  - 11
ER  -