Szx12 illb2 200 binocular microscope

The quantification of conidiospores was performed after three and five days of growth of A. nidulans colonies at 37 °C during constant illumination. Therefore, 5000 spores of the respective strains were point-inoculated on MM supplemented with 0.1% (v/v) pyridoxine, 5 mM uridine, and 0.1% (w/v) uracil. Spores were counted with Coulter Z2 particle counter (Beckman Coulter GmbH). For the analyses of sexual fruiting body formation, 5000 spores were either point-inoculated and incubated for seven days in darkness at 37 °C, or 30,000 spores were equally distributed on an agar plate and incubated for three, five, or seven days in darkness. Pictures of cleistothecia were taken with an Axiolab microscope (Carl Zeiss Microscopy, Oberkochen, Germany) and photomicrographs with the SZX12-ILLB2-200 binocular microscope (Olympus, Shinjuku, Japan).

Conidiospore numbers were determined with a Coulter Z2 particle counter (BECKMAN COULTER GMBH, Krefeld, Germany) or with a Thoma cell counting chamber (hemocytometer) (Marienfeld Superior). For quantifying cleistothecia, agar plugs of 5 mm² were cut out from plated using the larger side of a 200 μl pipette tip and cleistothecia were individualized on a fresh agar plate and counted with help of a binocular microscope SZX12-ILLB2-200 binocular microscope (Olympus).
ANOVA and t-test statistical analyses were conducted using standard deviations. Mutant samples were always compared to wildtype data for two-sample comparison through t-test.

Photomicrographs were obtained with an Axiolab microscope (Carl Zeiss Microscopy) and a SZX12-ILLB2-200 binocular microscope (Olympus). Fluorescence microscopy was performed with a Zeiss AxioObserver Z.1 inverted confocal microscope, equipped with Plan-Neofluar 63x/0.75 (air) and Plan-Apochromat 100x/1.4 oil objectives (Zeiss). The SlideBook 6.0 software (Intelligent Imaging Innovations) was used for picture processing.
Strains were grown in 8-well borosilicate cover glass system (Thermo Scientific) in 400 μl MM supplemented as mentioned above, when needed, or on glass slides covered with 1 ml solid MM supplemented as mentioned above, when needed, at 37°C or 30°C. GFP-signals were normalized against wildtype background signal to subtract fungal auto fluorescence. Nuclei were visualized by ectopic integration of ^pgpdA::rfp::h2A into the respective strains or through staining with 0.1% 4’,6’-diamidino-2phenylindole (DAPI).