W plan apochromat 40x 1.0 dic objective

Live cell imaging was performed as described previously (Prusicki et al, 2019 (link)). In short, up to six flower buds of 0.2–0.6 mm were carefully positioned in a petri plate filled with half-strength MS medium, pH 5.8 and solidified with 0.8% (w/v) agar. Time lapse was performed using an upright Zeiss LSM 880 confocal microscope with ZEN 2.3 SP1 software (Carl Zeiss AG, Oberkochen, Germany) and a W-plan Apochromat 40X/1.0 DIC objective (Carl Zeiss AG, Oberkochen, Germany). GFP and TagRPF were excited at λ = 488 nm and 561 nm, respectively, and detected between 498 and 560 nm and 520 and 650 nm, respectively. Auto-fluorescence was detected between 680 and 750 nm. With a time interval of 10 min, a series of six Z-stacks with 50 µm distance was acquired under a thermally controlled environment (21°C/30°C/34°C [± 0.15%]) in an incubation chamber. Due to sample movement, the Z-planes were manually selected using the review multi-dimensional data function of the software Metamorph version 7.8 and the XY movement was corrected using the Stack Reg plugin of Fiji.