TY - JOUR
T1 - A general method for bead-enhanced quantitation by flow cytometry
AU - Montes, Martin
AU - Jaensson, Elin A.
AU - Orozco, Aaron F.
AU - Lewis, Dorothy E.
AU - Corry, David B.
N1 - Funding Information:
We are grateful for the helpful suggestions of Dr. Farrah Kheradmand. Supported by NIH grants HL075243 and AI057696 (to D.B.C.), HDO46623 (to D.E.L. and A.F.O.), Fogarty Center training grant D43TW006569 (to M.M.) and the Sandler Program for Asthma Research (to D.B.C. and M.M.).
Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2006/12/20
Y1 - 2006/12/20
N2 - 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.
AB - 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.
KW - Apoptotic body
KW - Cell quantitation
KW - Eosinophil
KW - Flow cytometry
KW - T cell
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U2 - 10.1016/j.jim.2006.09.013
DO - 10.1016/j.jim.2006.09.013
M3 - Article
C2 - 17067632
AN - SCOPUS:33751343683
SN - 0022-1759
VL - 317
SP - 45
EP - 55
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
IS - 1-2
ER -