|
Fig. 1S4
alternate registration workflow for in situ pattern alignment.
A wild-type probe reference fish (PRF) is selected using iterative shape averaging following the workflow in Figure 1?figure supplement 3A,B. All wild-type and mutant embryos from the same age stained with the same probe are then registered to the PRF with rigid, similarity, and affine transformations in the green channel. To avoid deforming mutant gene expression patterns to more closely resemble wild-type gene expression patterns, separate mutant and wild-type reference fish are generated for the final non-linear B-spline transform. This is done by creating an average wild-type and an average mutant reference from the affine aligned images. All wild-type and mutant embryos are then registered to the appropriate reference with a B-spline transform.