Tcs sp2 scan head

roGFP2 measurements were done using an inverted microscope (Leica DMIRE2) equipped with a Leica TCS SP2 scan head (Leica Microsystems, Wetzlar, Germany). Conidia were prepared as described before [20 ]. Results were obtained by using the excitation wavelengths 395 (first track) and 488 (second track) as well as a 505–530 bandpass filter for collecting images. Z-stacks were displayed as average projections via the CLSM software. Further evaluation was done with the Image J program (v.1.44f; http://rsb.info.nih.gov/ij/) as it was shown before [33 (link)].
For measuring ROS levels, a Tecan Safire reader was used. Conidia were harvested and placed into a 96-well plate Microplate pureGradeTM 96 well PS, transp. Bottom, black, Brand GmbH & Co KG, Germany). After growth overnight (10⁵ conidia/ml) the detection mix was added and fluorescence was measured with 3 × 3 reads/well and excitation/emission as described above. The gain was set to 200.

roGFP2 measurements were done using an inverted microscope (Leica DMIRE2) equipped with a Leica TCS SP2 scan head (Leica Microsystems, Wetzlar, Germany). Conidia were prepared as described previously (Heller et al., 2012 (link)). Results were obtained by using the excitation wavelengths 395 (first track) and 488 (second track) as well as a 505–530 bandpass filter for collecting images. Z-stacks were displayed as average projections via the CLSM software. Further evaluation was done with the Image J program (v.1.44f¹) as it was shown before (Marschall et al., 2016b (link)).