A general method for bead-enhanced quantitation by flow cytometry

Martin Montes, Elin A. Jaensson, Aaron F. Orozco, Dorothy E. Lewis, David B. Corry

Research output: Contribution to journalArticle

34 Scopus citations

Abstract

Flow cytometry provides accurate relative cellular quantitation (percent abundance) of cells from diverse samples, but technical limitations of most flow cytometers preclude accurate absolute quantitation. Several quantitation standards are now commercially available which, when added to samples, permit absolute quantitation of CD4+ T cells. However, these reagents are limited by their cost, technical complexity, requirement for additional software and/or limited applicability. Moreover, few studies have validated the use of such reagents in complex biological samples, especially for quantitation of non-T cells. Here we show that addition to samples of known quantities of polystyrene fluorescence standardization beads permits accurate quantitation of CD4+ T cells from complex cell samples. This procedure, here termed single bead-enhanced cytofluorimetry (SBEC), was equally capable of enumerating eosinophils as well as subcellular fragments of apoptotic cells, moieties with very different optical and fluorescent characteristics. Relative to other proprietary products, SBEC is simple, inexpensive and requires no special software, suggesting that the method is suitable for the routine quantitation of most cells and other particles by flow cytometry.

Original languageEnglish (US)
Pages (from-to)45-55
Number of pages11
JournalJournal of Immunological Methods
Volume317
Issue number1-2
DOIs
StatePublished - Dec 20 2006

Keywords

  • Apoptotic body
  • Cell quantitation
  • Eosinophil
  • Flow cytometry
  • T cell

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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    Montes, M., Jaensson, E. A., Orozco, A. F., Lewis, D. E., & Corry, D. B. (2006). A general method for bead-enhanced quantitation by flow cytometry. Journal of Immunological Methods, 317(1-2), 45-55. https://doi.org/10.1016/j.jim.2006.09.013