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Deconvolution was performed with the "No Neighbour" method followed by a maximum projection using ImageJ (version 1.44 o).
Each slice measured 0.5 μm and stacks were reconstituted as a maximum projection using the MetaMorph software as described in detail [ 21].
From these stacks the four images closer to the middle section were selected and used for obtaining a maximum projection using the software Fiji.
To facilitate image analysis, 3D data sets were compressed into 2D images by using a maximum projection, using hms2Δ cells to determine threshold signal for expression.
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For KT-MT interaction analysis, z-stacks of optical sections were acquired at 0.3 µm intervals and images were deconvolved and reconstructed as maximum projections using AutoDeblur 9.3 software (AutoQuant Imaging Inc .. Measurements of fluorescence intensity were obtained from single optical sections of images acquired under identical exposure settings.
For the illustrations shown in Figure S5C, maximum projections using Image J were performed to generate 2D movies.
We remove these by first detecting the contour of the embryo in a maximum intensity projection using Otsu thresholding (Otsu, 1979) and marker-based watershed segmentation (Meyer, 1994), and then discarding all object calls that are outside the convex hull of the detected embryo contour.
Three dimensional reconstructions were rendered as maximum projection images using Volocity software (Perkin Elmer, Waltham, MA).
Stacks were collapsed into maximum intensity projections using Micro-Manager.
Stacks were collapsed into maximum intensity projections using Micro-Manager and stitched together into composite images using Photoshop.
Angiograms were obtained by generating maximum intensity projections using Paravision software (Bruker BioSpin GmbH).
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