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

The study presents a method for live imaging of leg regeneration in the crustacean Parhyale hawaiensis, capturing the entire process over up to 10 days at cellular resolution. This method addresses challenges such as minimizing photodamage and maintaining high spatial and temporal resolution. It involves mounting regenerating legs to minimize movement, using confocal microscopy for imaging, and employing computer-assisted cell tracking to determine cell lineages. The approach allows for the identification of cell fates and progenitors, potentially enabling the tracing of complete genealogies and characterization of molecular transitions during regeneration. This method overcomes previous limitations by avoiding long-term anesthesia and achieving continuous imaging with 20-minute intervals, providing a comprehensive view of the regeneration process.