Measuring lymphocyte proliferation, survival and differentiation using CFSE time-series data

Nat Protoc. 2007;2(9):2057-67. doi: 10.1038/nprot.2007.297.


Cellular proliferation is an essential feature of the adaptive immune response. The introduction of the division tracking dye carboxyfluorescein diacetate succinimidyl ester (CFSE) has made it possible to monitor the number of cell divisions during proliferation and to examine the relationship between proliferation and differentiation. Although qualitative examination of CFSE data may be useful, substantially more information about division and death rates can be extracted from quantitative CFSE time-series experiments. Quantitative methods can reveal in detail how lymphocyte proliferation and survival are regulated and altered by signals such as those received from co-stimulatory molecules, drugs and genetic polymorphisms. In this protocol, we present a detailed method for examining time-series data using graphical and computer-based procedures available to all experimenters.

MeSH terms

  • Cell Differentiation
  • Cell Division
  • Cell Proliferation
  • Cells, Cultured
  • Computer Simulation
  • Data Interpretation, Statistical
  • Flow Cytometry
  • Fluoresceins*
  • Fluorescent Dyes*
  • Lymphocyte Activation*
  • Lymphocytes / cytology
  • Lymphocytes / immunology*
  • Models, Biological
  • Software*
  • Staining and Labeling
  • Succinimides*


  • 5-(6)-carboxyfluorescein diacetate succinimidyl ester
  • Fluoresceins
  • Fluorescent Dyes
  • Succinimides