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Movies of the image stacks were generated with the frame rate of 25 fps.
3D projections of the image stacks were generated with the orthogonal perspective for the representative images shown in figures.
Image stacks were generated using a Nikon A1R confocal microscope, and used to produce single, projected images.
To determine CenH3CID fluorescence signal intensities in S2 cells, z stacks were generated of 8 to 10 optical sections in the green channel using maximum intensity projection mode.
For CenH3CID intensity measurements of stage 4 embryos, z stacks were generated of 3 to 4 optical sections in the green channel using maximum intensity projection mode.
Z-projections of cell image stacks were generated using ImageJ.
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Z-stacks were generated by obtaining optical sections through the samples at 0.35 µm intervals along the z axis.
3D renderings of confocal z-stacks were generated and analyzed using Imaris 6.4 3D image visualization and analysis software (Bitplane Inc., Saint Paul, MN, USA).
The Z-stacks were generated in 0.45 µm increments and 3-D reconstructions were created by computer using Velocity software (Impro Vision Inc., Lexington, MA).
To exclude the loss of FISH signals, three-dimensional z-stacks were generated.
Three-dimensional z-stacks were generated, colours were separately recorded and digitally processed.
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