Elucidation of seventeen human peripheral blood B‐cell subsets and quantification of the tetanus response using a density‐based method for the automated …

Y Qian, C Wei, F Eun‐Hyung Lee… - Cytometry Part B …, 2010 - Wiley Online Library
Y Qian, C Wei, F Eun‐Hyung Lee, J Campbell, J Halliley, JA Lee, J Cai, YM Kong, E Sadat…
Cytometry Part B: Clinical Cytometry, 2010Wiley Online Library
Background: 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. Methods: To address
this challenge, we developed a novel program, FLOCK (FLOw Clustering without K), that …
Background
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.
Methods
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.
Results
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.
Conclusions
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. © 2010 International Clinical Cytometry Society
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