CALIPERS: Cell cycle-aware live imaging for phenotyping experiments and regeneration studies

Summary

Cell-cycle progression is a major source of variability in live-cell phenotyping, yet imaging workflows still lack a general way to account for it alongside structural and functional readouts. Here we show CALIPERS (Cell-cycle-Aware Live-cell Imaging for Phenotyping Experiments and Regeneration Studies), an integrated framework that pairs a spectrally redesigned FUCCI reporter with continuous cell-cycle inference and flexible delivery strategies in human cells and stem-cell models. In epithelial cells, CALIPERS simultaneously images actin, tubulin, or calcium dynamics, phase-locks migration and proliferation, triggers mitosis-aware smart microscopy, and recovers fast calcium signals. In induced pluripotent stem-cell workflows, lentiviral and safe-harbor versions span pluripotent, lineage-restricted, and cardiac models. In cardiac organoids, CALIPERS tracks cell-cycle exit, tissue compaction, and calcium onset, and distinguishes productive proliferation from multinucleation and endoreplication. By making cell-cycle state an explicit, measurable variable, CALIPERS helps triage candidate regenerative interventions across diverse live-imaging assays. © 2026. The Author(s).

Authors Di Sante M, Pezzotti M, Zimmermann J, Enrico A, Deschamps J, Balmas E, Becca S, Torchia E, Ballio G, Solito S, Sarchi M, Reali A, Bertero A, Jug F, Pasqualini FS
Journal Nature communications
Publication Date 2026 Aug 1;17(1)
PubMed 42669673
PubMed Central PMC13526800
DOI 10.1038/s41467-026-76144-9

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