Zebrafish Caudal Fin Regeneration

Regeneration following amputation was quantified at various timepoints, with a different batch of fish used for each respective timepoint. Zebrafish were euthanized in 50 μg/mL benzocaine and fully mineralized bone matrix was visualized using Alizarin red, and newly depositing bone matrix was visualized using calcein green [20 (link)]. Regenerating caudal fins were double-stained with Alizarin red (74 μM Alizarin red S, 5 mM HEPES (Promega, USA) and calcein (40 μM calcein green) made up in 1× Danieau, as previously described [63 (link)]. The region of amputation was imaged under UV excitation through the GFP and RFP filter for calcein and Alizarin red visualization, respectively.