Identification of B cells through negative gating-An example of the MIFlowCyt standard applied.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
Darren BlimkieTobias Kollmann

Abstract

Polychromatic flow cytometric analysis takes advantage of the increasing number of available fluorophores to positively identify and simultaneously assess multiple parameters in the same cell (1). Additional parameters may be analyzed through negative identification (i.e., through exclusion of particular stains or antibodies employed). In this report, we tested whether such negative-gating strategy would identify human B lymphocytes in innate immune phenotyping studies. To this end, B cells were identified as the negatively-stained subpopulation from the CD123 vs. CD11c plot of the CD14(neg-low), MHC II(high) human peripheral blood mononuclear cells. To test the specificity of this negative gating approach, we confirmed that negatively gated B cells indeed expressed CD19, the bona fide marker for human B cells. However, a small number of unidentified cells were contained in the negatively-gated B cells. Furthermore, a small percentage cells expressing markers used to identify monocytes and myeloid dendritic cells (mDC) coexpressed CD19. This identifies such negative B-cell gating approach as potentially problematic. When applied to the analysis of Toll-like receptors (TLR) stimulation experiments, we were however able to interp...Continue Reading

References

Mar 17, 2006·European Journal of Immunology·Claudia R Ruprecht, Antonio Lanzavecchia
Jun 2, 2006·Molecular Systems Biology·John Quackenbush
Jun 21, 2006·Nature Immunology·Leonore A HerzenbergDavid R Parks
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Citations

Apr 16, 2014·Journal of Acquired Immune Deficiency Syndromes : JAIDS·Brian A ReikieTobias R Kollmann
Feb 1, 2011·Journal of Immunological Methods·Darren BlimkieTobias R Kollmann
Aug 20, 2010·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Mario Roederer, Attila Tárnok
Apr 21, 2012·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Attila Tárnok
Aug 24, 2012·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Attila Tárnok
Apr 19, 2017·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Kinga K SmolenTobias R Kollmann
Aug 20, 2014·The Journal of Immunology : Official Journal of the American Association of Immunologists·Kinga K SmolenTobias R Kollmann

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