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A novel statistical analysis and interpretation of flow cytometry data.

Banks HT, Kapraun DF, Thompson WC, Peligero C, Argilaguet J, Meyerhans A - J Biol Dyn (2013)

Bottom Line: A recently developed class of models incorporating the cyton model of population generation structure into a conservation-based model of intracellular label dynamics is reviewed.Statistical aspects of the data collection process are quantified and incorporated into a parameter estimation scheme.This scheme is then applied to experimental data for PHA-stimulated CD4+T and CD8+T cells collected from two healthy donors.

View Article: PubMed Central - PubMed

Affiliation: Center for Research in Scientific Computation and Center for Quantitative Sciences in Biomedicine, North Carolina State University, Raleigh, NC 27695-8212, USA. htbanks@ncsu.edu

ABSTRACT
A recently developed class of models incorporating the cyton model of population generation structure into a conservation-based model of intracellular label dynamics is reviewed. Statistical aspects of the data collection process are quantified and incorporated into a parameter estimation scheme. This scheme is then applied to experimental data for PHA-stimulated CD4+T and CD8+T cells collected from two healthy donors. This novel mathematical and statistical framework is shown to form the basis for accurate, meaningful analysis of cellular behaviour for a population of cells labelled with the dye carboxyfluorescein succinimidyl ester and stimulated to divide.

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Measured autofluorescence distributions in comparison to fitted lognormal curves for Donor 1 (left) and Donor 2 (right) for CD8T cells. The least-squares fit to data is visually better than the fit obtained by the method of moments (MM), though the difference is small.
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Figure 4: Measured autofluorescence distributions in comparison to fitted lognormal curves for Donor 1 (left) and Donor 2 (right) for CD8T cells. The least-squares fit to data is visually better than the fit obtained by the method of moments (MM), though the difference is small.

Mentions: To test the assumption of lognormality, a portion of PBMCs from each donor were set aside and stimulated with PHA but never labelled with CFSE. Thus the measured fluorescence distribution of these cells (represented in histogram form) can be used to approximate the density function p(t,ζ) representing the actual population distribution of autofluorescence. This autofluorescence data are depicted for the two donors and cell types in Figures 3 and 4. To assess the lognormal approximation, we used two parameter estimation schemes to construct lognormal density functions from the autofluorescence data. The first method is the method of moments, in which the exact mean and variance of the measured cells was computed and a lognormal curve was constructed with the same mean and variance. (In the figures, the resulting lognormal density function has been scaled by the number of cells in the data to facilitate comparison.) The second method is to use least squares to estimate the scale, mean, and variance of a lognormal density function.


A novel statistical analysis and interpretation of flow cytometry data.

Banks HT, Kapraun DF, Thompson WC, Peligero C, Argilaguet J, Meyerhans A - J Biol Dyn (2013)

Measured autofluorescence distributions in comparison to fitted lognormal curves for Donor 1 (left) and Donor 2 (right) for CD8T cells. The least-squares fit to data is visually better than the fit obtained by the method of moments (MM), though the difference is small.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3753657&req=5

Figure 4: Measured autofluorescence distributions in comparison to fitted lognormal curves for Donor 1 (left) and Donor 2 (right) for CD8T cells. The least-squares fit to data is visually better than the fit obtained by the method of moments (MM), though the difference is small.
Mentions: To test the assumption of lognormality, a portion of PBMCs from each donor were set aside and stimulated with PHA but never labelled with CFSE. Thus the measured fluorescence distribution of these cells (represented in histogram form) can be used to approximate the density function p(t,ζ) representing the actual population distribution of autofluorescence. This autofluorescence data are depicted for the two donors and cell types in Figures 3 and 4. To assess the lognormal approximation, we used two parameter estimation schemes to construct lognormal density functions from the autofluorescence data. The first method is the method of moments, in which the exact mean and variance of the measured cells was computed and a lognormal curve was constructed with the same mean and variance. (In the figures, the resulting lognormal density function has been scaled by the number of cells in the data to facilitate comparison.) The second method is to use least squares to estimate the scale, mean, and variance of a lognormal density function.

Bottom Line: A recently developed class of models incorporating the cyton model of population generation structure into a conservation-based model of intracellular label dynamics is reviewed.Statistical aspects of the data collection process are quantified and incorporated into a parameter estimation scheme.This scheme is then applied to experimental data for PHA-stimulated CD4+T and CD8+T cells collected from two healthy donors.

View Article: PubMed Central - PubMed

Affiliation: Center for Research in Scientific Computation and Center for Quantitative Sciences in Biomedicine, North Carolina State University, Raleigh, NC 27695-8212, USA. htbanks@ncsu.edu

ABSTRACT
A recently developed class of models incorporating the cyton model of population generation structure into a conservation-based model of intracellular label dynamics is reviewed. Statistical aspects of the data collection process are quantified and incorporated into a parameter estimation scheme. This scheme is then applied to experimental data for PHA-stimulated CD4+T and CD8+T cells collected from two healthy donors. This novel mathematical and statistical framework is shown to form the basis for accurate, meaningful analysis of cellular behaviour for a population of cells labelled with the dye carboxyfluorescein succinimidyl ester and stimulated to divide.

Show MeSH