Moises Di Sante, Melissa Pezzotti, Julius Zimmermann, Alessandro Enrico, J.R. Deschamps, Elisa Balmas, Silvia Becca, Eloisa Torchia, Giulia Ballio, Samantha Solito, Martina Sarchi, Alessandro Reali, Alessandro Bertero, Florian Jug, Francesco S. Pasqualini
Abstract Cell cycle progression, migration, and proliferation shape development and regeneration, but simultaneous live-cell imaging remains challenging as conventional fluorescent cell cycle indicators (FUCCI) monopolize the green and red channels used by most structural and functional biosensors. To overcome this, we integrated a spectrally re-engineered FUCCI variant, open-source analysis software, and four-color human stem cell reporter lines into CALIPERS: a method for Cell-cycle-Aware Live-cell Imaging in Phenotyping and Regeneration Studies.