2020, Frawley, Andrew T., Wycisk, Virginia, Xiong, Yaoyao, Galiani, Silvia, Sezgin, Erdinc, Urbančič, Iztok, Vargas Jentzsch, Andreas, Leslie, Kathryn G., Eggeling, Christian, Anderson, Harry L.

Dyads consisting of a photochromic switch covalently linked to a fluorescent dye allow the emission from the dye to be controlled by reversible photoisomerization of the switch; one form of the switch quenches fluorescence by accepting energy from the dye. Here we investigate the use of dyads of this type for super-resolution imaging of lipid bilayers. Giant unilamellar vesicles stained with the dyads were imaged with about a two-fold resolution-enhancement compared with conventional confocal microscopy. This was achieved by exciting the fluorophore at 594 nm, using a switch activated by violet and red light (405/640 nm). This journal is © The Royal Society of Chemistry.

2019, Karras, Christian, Smedh, Maria, Förster, Ronny, Deschout, Hendrik, Fernandez-Rodriguez, Julia, Heintzmann, Rainer

The impact of the different reconstruction parameters in super-resolution structured illumination microscopy (SIM) on image artifacts is carefully analyzed. These parameters comprise the Wiener filter parameter, an apodization function, zero-frequency suppression and modifications of the optical transfer function. A detailed investigation of the reconstructed image spectrum is concluded to be suitable for identifying artifacts. For this purpose, two samples, an artificial test slide and a more realistic biological system, were used to characterize the artifact classes and their correlation with the image spectra as well as the reconstruction parameters. In addition, a guideline for efficient parameter optimization is suggested and the implementation of the parameters in selected up-to-date processing packages (proprietary and open-source) is depicted. © 2018 The Authors