Plan neofluor z objective

Transgenic larvae were pre-screened for fluorescence using a zoomscope (EMS3/SyCoP3; Zeiss; Plan-NeoFluor Z objective). For multi-day imaging experiments, larvae were anesthetized and mounted in a Z-wedgi device [43 (link),48 (link)] where they were oriented such that the hindbrain was fully visible. Z-series images (5 μm slices) of the hindbrain were acquired on a spinning disk confocal microscope (CSU-X; Yokogawa) with a confocal scan head on a Zeiss Observer Z.1 inverted microscope, Plan-Apochromat NA 0.8/20x objective, and a Photometrics Evolve EMCCD camera. Between imaging sessions larvae were kept in E3-MB with PTU in individual wells of 24- or 48-well plates. Neutrophil-fungal interactions were imaged using an inverted epifluorescence microscope (Nikon Eclipse TE3000) with a Nikon Plan Fluor 20x/0.50 objective, motorized stage (Ludl Electronic Products) and Prime BSI Express camera (Teledyne Photometrics). Environmental controls were set to 37°C with 5% CO₂. Images were acquired every 3 min for 12 h. Imaging of A. fumigatus stained with CFW was performed using an upright Zeiss Imager.Z2 LSM 800 laser scanning confocal microscope with Airyscan detection and a Plan-Apochromat 20x /0.8 objective. A single z plane image was acquired for each hypha. Images were captured using identical laser and exposure settings for each condition.

Transgenic larvae were pre-screened for fluorescence using a zoomscope (EMS3/SyCoP3; Zeiss; Plan-NeoFluor Z objective). For multi-day imaging experiments, larvae were anesthetized and mounted in a Z-wedgi device [39 , 44 (link)] where they were oriented such that the hindbrain was fully visible. Z-series images (5 μm slices) of the hindbrain were acquired on a spinning disk confocal microscope (CSU-X; Yokogawa) with a confocal scanhead on a Zeiss Observer Z.1 inverted microscope, Plan-Apochromat NA 0.8/20x objective, and a Photometrics Evolve EMCCD camera. Between imaging sessions larvae were kept in E3-MB with PTU in individual wells of 24- or 48-well plates. Neutrophil-fungal interactions were imaged using an inverted epifluorescence microscope (Nikon Eclipse TE3000) with a Nikon Plan Fluor 20x/0.50 objective, motorized stage (Ludl Electronic Products) and Prime BSI Express camera (Teledyne Photometrics). Environmental controls were set to 37°C with 5% CO₂. Images were acquired every 3 min for 12 h. Imaging of A. fumigatus stained with CFW was performed using an upright Zeiss Imager.Z2 LSM 800 laser scanning confocal microscope with Airyscan detection and a Plan-Apochromat 20x /0.8 objective. A single z plane image was acquired for each hypha. Images were captured using identical laser and exposure settings for each condition.