Registration

We developed a registration algorithm based on iterative optical flow estimation¹ that estimates motion patch-wise and combines smoothness regularization, signal-to-noise weighting of gradients, and multi-resolution registration to favor convergence to an optimal tissue alignment for each time point. This achieved high-quality alignment, and most cells were accurately registered throughout the experiments. To evaluate registration accuracy, we employed fiduciary landmarks provided by a sparse transgenic line Tg(phox2bb:eGFP)², used solely for assessment. Although at times the rapid contractions of gastrointestinal tissues posed challenges, most cells were well-registered for almost all time points. Finally, a registration reliability map is applied to exclude specific subregions at time points with uncertain registration accuracy.

Registration alogrithm

Workflow for WBI data registration utilizing multi-scale iterative estimation of local motion fields, with example results demonstrating temporal alignment of the tissue.

Registration results

References

• 1. Determining Optical Flow, , 1981, Horn, Berthold K. and Schunck, Brian G.
• 2. Specification of epibranchial placodes in zebrafish, Development, 2007, Nechiporuk, Alexei and Linbo, Tor and Poss, Kenneth D. and Raible, David W.