Photometrix cool snaphq camera

The initiation of fruiting body development was quantified after 3 days of incubation on BMM-covered microscope slides. Ascogonia and protoperithecia were counted separately within an area of 0.5 cm² located 1 cm behind the growth front. An Axio Imager M1 (Zeiss) with a SpectraX LED lamp (Lumencor) and a Photometrix Cool SnapHQ camera (Roper Scientific, Martinsried, Germany) was used for quantification. Experiments were repeated for three biological replicats per strain. For assessment of fruiting body formation, images of strains were taken after 7 days of incubation in Petri dishes on solid BMM with a Zeiss Stemi 2000-C binocular, using an AxioCam ERc5s with the software ZEN 2 core (version 2.5, Zeiss, Jena, Germany).

To investigate the subcellular localization of RTT109-EGFP fusion proteins, the strains were cultivated on glass slides along with strains expressing H3-mRFP fusion proteins, as previously described (Schmidt et al. 2020 (link)). Heterokaryotic strains can therefore express both fluorescence-labeled proteins (Engh et al. 2007 (link)). Subsequently, light and fluorescence microscopy were performed using an AxioImager microscope (Zeiss), equipped with a Photometrix Cool SnapHQ camera (Roper Scientific). EGFP fluorescence was detected using a Chroma filter set 41017 (HQ470/40, HQ525/50, Q495lp), while mRFP fluorescence was detected using set 49008 (EG560/40x, ET630/75m, T585lp). The acquired images were further processed using MetaMorph software (Molecular Devices).

For microscopy of mycelia undergoing sexual development, S. macrospora strains were grown on glass slides with a thin layer of corn meal extract or corn meal medium solidified with 0.8% agar as described [117 (link), 118 (link)]. Fluorescence and light microscopic investigations were carried out with an AxioImager microscope (Zeiss, Jena, Germany). Fluorescence was studied using Chroma (Bellows Falls, VT, USA) filter set 41017 (HQ470/40, HQ525/50, Q495lp) for detection of EGFP, set 49008 (EG560/40x, ET630/75 m, T585lp) for the detection of tdTomato, and set 31000v2 (D350/50, D460/50, 400dclp) for the detection of DAPI and mKalama1. Images were captured with a Photometrix Cool SnapHQ camera (Roper Scientific) and MetaMorph (Universal Imaging). Recorded images were edited with MetaMorph and Adobe Photoshop CS4.