Automated gating of flow cytometry data via robust model-based clustering

Cytometry A. 2008 Apr;73(4):321-32. doi: 10.1002/cyto.a.20531.


The capability of flow cytometry to offer rapid quantification of multidimensional characteristics for millions of cells has made this technology indispensable for health research, medical diagnosis, and treatment. However, the lack of statistical and bioinformatics tools to parallel recent high-throughput technological advancements has hindered this technology from reaching its full potential. We propose a flexible statistical model-based clustering approach for identifying cell populations in flow cytometry data based on t-mixture models with a Box-Cox transformation. This approach generalizes the popular Gaussian mixture models to account for outliers and allow for nonelliptical clusters. We describe an Expectation-Maximization (EM) algorithm to simultaneously handle parameter estimation and transformation selection. Using two publicly available datasets, we demonstrate that our proposed methodology provides enough flexibility and robustness to mimic manual gating results performed by an expert researcher. In addition, we present results from a simulation study, which show that this new clustering framework gives better results in terms of robustness to model misspecification and estimation of the number of clusters, compared to the popular mixture models. The proposed clustering methodology is well adapted to automated analysis of flow cytometry data. It tends to give more reproducible results, and helps reduce the significant subjectivity and human time cost encountered in manual gating analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Antibodies, Monoclonal / pharmacology
  • Antibodies, Monoclonal, Murine-Derived
  • Cluster Analysis*
  • Computational Biology / methods
  • Computer Simulation
  • Drug Screening Assays, Antitumor*
  • Flow Cytometry / methods*
  • Humans
  • Likelihood Functions
  • Models, Statistical
  • Models, Theoretical
  • Multivariate Analysis
  • Normal Distribution
  • Rituximab


  • Antibodies, Monoclonal
  • Antibodies, Monoclonal, Murine-Derived
  • Rituximab