Roots were stained by propidium iodide (PI; Sigma-Aldrich) at final concentration of 10μg/ml. Embryos were fixed and stained by 2.2% Renaissance RS2200 (Renaissance Chemicals Ltd) in PBS buffer at pH 6.9 containing 4% paraformaldehyde, 5% glycerol, 4.2% dimethyl sulfoxide (DMSO) or stained by PI after fixation as previously described6 (link),38 (link).
Confocal imaging was performed with Zeiss LSM700/800 (observation of lateral root) or Leica SP5/II (observation of primary root and embryo) as previously described38 (link). Live imaging was performed with LSM700 as described before39 (link),40 (link). For the 3D imaging of pSOK1-SOK1-YFP and pSOK1-GFP root, Zeiss LSM780 with 2 photon laser (960-990nm) and 500-550nm/575-610nm band-pass filter were used. Analysis of confocal images (3D reconstruction, 3D segmentation) was performed by MorphoGraphX 41 (link).
Confocal imaging was performed with Zeiss LSM700/800 (observation of lateral root) or Leica SP5/II (observation of primary root and embryo) as previously described38 (link). Live imaging was performed with LSM700 as described before39 (link),40 (link). For the 3D imaging of pSOK1-SOK1-YFP and pSOK1-GFP root, Zeiss LSM780 with 2 photon laser (960-990nm) and 500-550nm/575-610nm band-pass filter were used. Analysis of confocal images (3D reconstruction, 3D segmentation) was performed by MorphoGraphX 41 (link).
