Regita 1300 qimaging camera

Analysis of dry seed morphology parameters was performed in three biological replicates, each with at least 20 seeds collected from four to five spikes of different plants. Dry kernels were peeled off, weighed with an analytical scale (Sartorius, Göttingen, Germany), and photographed using a SZX16 binocular microscope (Olympus, Tokyo, Japan) bonded with a Regita 1300 QImaging camera and QCapture ×64 software (Olympus). Seed length and width were measured using ImageJ calibrated with internal size control. Seeds from 4 to 48 DAP and dry seeds were peeled off and cut with a razor blade along the longitudinal and transverse axis. At least 20 individual seeds were photographed as described above using a binocular microscope. Hulled seeds that possessed awns were photographed with a D5600 (Nikon, Tokyo, Japan) digital camera equipped with an 80 mm Nikkor objective. All photo-matrix were composed of separately taken photos of individual seeds and merged in Adobe Photoshop CS5 (Adobe Inc., San Jose, CA, USA).

Seeds from 4 to 48 DAP and dry seeds were peeled off and cut with a razor blade along the longitudinal and transverse axis. At least 20 individual seeds bulked from four to five spikes of different plants were stained in 0.1% (w/v) Evans blue (314-13-6, Sigma-Aldrich, St. Louis, MO, USA) for 2 min. Stained sections were washed twice for 10 min with distilled water [31 (link)]. Transverse and sagittal sections of samples were analyzed with an SZX16 binocular microscope (Olympus). Images were captured with a Regita 1300 QImaging camera and QCapture ×64 software (Olympus) using the same settings and proceeded in Adobe Photoshop CS5 (Adobe Inc.).