This set of images corresponds to wholemount immunolocalized Col0 untreated root tips obtained using an anti-tubulin antibody (B-5-1-2 mouse monoclonal anti-tubulin, Sigma-Aldrich), an anti-Histone H3 (Phospho S10) antibody (rabbit polyclonal anti-H3S10ph antibody) and stained with DAPI. Root tips Z-stacks were acquired using an SP8 confocal laser microscope (Leica Microsystems). For simultaneous detection of DAPI, microtubules and H3S10ph signals, samples were excited respectively at 405 nm, 488 nm or 561 nm, with an emission band of 420-446 nm, 495-540 nm or 570-620 nm, using PMT detectors for both DAPI and H3S10ph, and a hybrid detector for microtubules. All 3D stacks were acquired as 12 bits images to get a high dynamic range of the H3S10ph signal, ensuring that no pixel was saturated in this channel. All 3D stacks were acquired with using the same zoom, the same voxel size (350 µm x 350 µm x 350 µm) and the same settings and PMT level for the H3S10ph channel.