Quantifying Immune Cell Dynamics in Zebrafish Tail Regeneration

4 dpf Tg(mpx:GFP/mpeg:mCherry) embryos were anesthetized with tricaine and tails were transected distal to the notochord using a scalpel blade. Embryos were immediately mounted for confocal imaging and imaged from ∼30 min post wounding for 7 h every minute. Particles were detected and tracks were analyzed using the Trackmate plugin from Fiji (Tinevez et al., 2017 (link)). The linear motion algorithm was used with a maximum allowed gap of four timeframes and a maximum allowed radius of 50 µm. The wound area was defined as 200 μm and closer from the edge of the tail in the first timeframe. The mean speed of tracks outside the wound area was determined by dividing the total distance by the total time of the track. The meandering index of tracks outside the wound area was determined by dividing the net distance (distance between the first and last point of track) by the total distance of the track. Wound persistence was determined for each track that entered the wound area by dividing the number of timeframes spent inside the wound area by the number of timeframes left till the end of the movie.