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  - 
TY  - JOUR
A1  - Proppert, Sven
A1  - Wolter, Steve
A1  - Holm, Thorge
A1  - Klein, Theresa
A1  - van de Linde, Sebastian
A1  - Sauer, Markus
T1  - Cubic B-spline calibration for 3D super-resolution measurements using astigmatic imaging
JF  - Optics Express
N2  - In recent years three-dimensional (3D) super-resolution fluorescence imaging by single-molecule localization (localization microscopy) has gained considerable interest because of its simple implementation and high optical resolution. Astigmatic and biplane imaging are experimentally simple methods to engineer a 3D-specific point spread function (PSF), but existing evaluation methods have proven problematic in practical application. Here we introduce the use of cubic B-splines to model the relationship of axial position and PSF width in the above mentioned approaches and compare the performance with existing methods. We show that cubic B-splines are the first method that can combine precision, accuracy and simplicity.
KW  - three-dimensional microscopy
KW  - fluorescence microscopy
KW  - medical and biological imaging
KW  - superresolution
Y1  - 2014
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-119730
SN  - 1094-4087
VL  - 22
IS  - 9
ER  -