Ec plan neofluar 10 0.30 ph 1 objective

For fluorescence images, fresh tissue biopsies were snap-frozen in Tissue-Tek (VWR) immediately after removal. Serial 12 μm cryosections were prepared with a microtome and fixed with ice-cold acetone for 10 minutes. Sections were mounted using Mowiol 4–88 (Roth) and phase contrast and fluorescence micrographs were taken on an ApoTome.2 (Zeiss) equipped with a Plan-Apochromat 63×/1.40 Oil DIC M27 objective and an EC Plan-Neofluar 10×/0.30 Ph 1 objective.
For histological stainings, fresh tissue biopsies were fixed over night using 4% formaldehyde and embedded in paraffin. 5 μm sections were prepared with a microtome and stained with hematoxylin-eosin. Micrographs were taken on an Eclipse 80i microscope (Nikon).

Live cell analyses and visualization of expressed fluorescent eGFP signals were performed 24 h after treatment with Fuse-It-mRNA or Lipofectamine^® 2000 at 37 °C and 5% CO₂ using an inverse confocal laser scanning microscope (cLSM 710, Carl Zeiss Jena, Germany). All images showed a representative overview of the center of the substrate and were recorded with an EC “Plan-Neofluar” 10×/0.30 Ph1 objective (Carl Zeiss Jena, Germany). For all experiments, eGFP-fluorescence was excited by a 488 nm argon laser. Emission was detected in a range of 500–550 nm. Throughout all experiments, the microscope settings were kept unchanged to allow for the best data comparability.

Flowers were harvested from 10-week-old Arabidopsis plants grown under long-day conditions. Seeds were air-dried for 2 h and then slowly rehydrated in a humid chamber for 45 min at 30°C. Flowers were gently dipped into working solution [17% (w/v) sucrose with 2 μg mL⁻¹ of FDA (fluorescein diacetate) (Heslop-Harrison and Heslop-Harrison, 1970 (link))] to release the pollen. After 15 min, the pollen grains were observed in the same media under a fluorescence microscope (Axio Observer Z1 inverted microscope with AxioCamMR3, from Zeiss) with EC Plan-Neofluar 10 × /0.30 Ph1 objective (Zeiss) and 38 HE Green Fluorescence Reflector (Zeiss). For image acquisition and analysis, AxioVision Rev 4.8 software was used. Fluorescent pollen grains were counted as viable.