Introduction

Advances in multiparameter flow cytometry (FCM) now allow for the independent detection of larger numbers of fluorochromes on individual cells, generating data with increasingly higher dimensionality. The increased complexity of these data has made it difficult to identify cell populations from high-dimensional FCM data using traditional manual gating strategies based on single-color or two-color displays.To address this challenge, we developed a novel program, FLOCK (FLOw Clustering without K), that uses a density-based clustering approach to algorithmically identify biologically relevant cell populations from multiple samples in an unbiased fashion, thereby eliminating operator-dependent variability.FLOCK was used to objectively identify seventeen distinct B-cell subsets in a human peripheral blood sample and to identify and quantify novel plasmablast subsets responding transiently to tetanus and other vaccinations in peripheral blood. FLOCK has been implemented in the publically available Immunology Database and Analysis Portal-ImmPort (http://www.immport.org)-for open use by the immunology research community.FLOCK is able to identify cell subsets in experiments that use multiparameter FCM through an objective, automated computational approach. The use of algorithms like FLOCK for FCM data analysis obviates the need for subjective and labor-intensive manual gating to identify and quantify cell subsets. Novel populations identified by these computational approaches can serve as hypotheses for further experimental study.

Publications

  1. Elucidation of seventeen human peripheral blood B-cell subsets and quantification of the tetanus response using a density-based method for the automated identification of cell populations in multidimensional flow cytometry data.
    Cite this
    Qian Y, Wei C, Eun-Hyung Lee F, Campbell J, Halliley J, Lee JA, Cai J, Kong YM, Sadat E, Thomson E, Dunn P, Seegmiller AC, Karandikar NJ, Tipton CM, Mosmann T, Sanz I, Scheuermann RH, 2010-01-01 - Cytometry. Part B, Clinical cytometry

Credits

  1. Yu Qian
    Developer

    Department of Pathology, University of Texas Southwestern Medical Center, United States of America

  2. Chungwen Wei
    Developer

  3. F Eun-Hyung Lee
    Developer

  4. John Campbell
    Developer

  5. Jessica Halliley
    Developer

  6. Jamie A Lee
    Developer

  7. Jennifer Cai
    Developer

  8. Y Megan Kong
    Developer

  9. Eva Sadat
    Developer

  10. Elizabeth Thomson
    Developer

  11. Patrick Dunn
    Developer

  12. Adam C Seegmiller
    Developer

  13. Nitin J Karandikar
    Developer

  14. Christopher M Tipton
    Developer

  15. Tim Mosmann
    Developer

  16. Iñaki Sanz
    Developer

  17. Richard H Scheuermann
    Investigator

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Summary
AccessionBT000112
Tool TypeApplication
Category
PlatformsLinux/Unix
TechnologiesC
User InterfaceTerminal Command Line
Download Count0
Submitted ByRichard H Scheuermann