Long-Term Live Imaging, Cell Identification and Cell Tracking in Regenerating Crustacean Legs

The study presents a method for high-resolution live imaging of leg regeneration in the crustacean Parhyale hawaiensis, capturing the entire process over up to 10 days at cellular resolution. The method involves mounting and imaging regenerating legs to achieve high spatial and temporal resolution while minimizing photodamage, fixing and staining the regenerated legs to identify cell fates, and using computer-assisted cell tracking to determine cell lineages and progenitors. This approach is optimized to limit light exposure and maximize tracking efficiency, potentially allowing for the description of cell lineages for every regenerated cell type in the limb.