Quantitative Cell Cycle Analysis Based on an Endogenous All-in-One Reporter for Cell Tracking and Classification

Cell Rep. 2017 May 30;19(9):1953-1966. doi: 10.1016/j.celrep.2017.05.022.


Cell cycle kinetics are crucial to cell fate decisions. Although live imaging has provided extensive insights into this relationship at the single-cell level, the limited number of fluorescent markers that can be used in a single experiment has hindered efforts to link the dynamics of individual proteins responsible for decision making directly to cell cycle progression. Here, we present fluorescently tagged endogenous proliferating cell nuclear antigen (PCNA) as an all-in-one cell cycle reporter that allows simultaneous analysis of cell cycle progression, including the transition into quiescence, and the dynamics of individual fate determinants. We also provide an image analysis pipeline for automated segmentation, tracking, and classification of all cell cycle phases. Combining the all-in-one reporter with labeled endogenous cyclin D1 and p21 as prime examples of cell-cycle-regulated fate determinants, we show how cell cycle and quantitative protein dynamics can be simultaneously extracted to gain insights into G1 phase regulation and responses to perturbations.

Keywords: G1 phase regulation; automated image analysis; cell cycle; cell cycle reporter; cell fate decisions; cyclin D1; cyclin oscillations; p21; quantitative single cell imaging; quiescence.

MeSH terms

  • Animals
  • Cell Cycle*
  • Cell Survival
  • Cell Tracking / methods*
  • Cyclin-Dependent Kinase Inhibitor p21 / metabolism
  • Cyclins / metabolism
  • DNA Damage
  • G1 Phase
  • Genes, Reporter*
  • Humans
  • Kinetics
  • Mice
  • Proliferating Cell Nuclear Antigen / metabolism


  • Cyclin-Dependent Kinase Inhibitor p21
  • Cyclins
  • Proliferating Cell Nuclear Antigen